KINDAI UNIVERSITY


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NISHIDA Shozo

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FacultyDepartment of Pharmacy / Graduate School of Medicine
PositionProfessor
Degree
Commentator Guidehttps://www.kindai.ac.jp/meikan/758-nishida-shouzou.html
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Last Updated :2020/09/04

Education and Career

Education

  •  - 1988 , Kindai University, Graduate School of Medical Sciences
  • Kindai University, Faculty of Pharmacy
  •  - 1988 , Kinki University, Graduate School, Division of Medicine
  • Kinki University, Faculty of Pharmaceutical Science

Research Activities

Research Areas

  • Life sciences, Pharmacology
  • Life sciences, Experimental pathology
  • Life sciences, Pharmacology
  • Life sciences, Pharmaceuticals - health and biochemistry

Research Interests

  • Experimental Pathology, Biochemistry, Pharmacotherapy

Published Papers

  • Overexpression of HIF-1α contributes to melphalan resistance in multiple myeloma cells by activation of ERK1/2, Akt, and NF-κB., Masanobu Tsubaki, Tomoya Takeda, Yoshika Tomonari, Yu-Ichi Koumoto, Motohiro Imano, Takao Satou, Shozo Nishida, Laboratory investigation; a journal of technical methods and pathology, Laboratory investigation; a journal of technical methods and pathology, 99(1), 72 - 84, Jan. 2019 , Refereed
    Summary:Multiple myeloma (MM) commonly displays multidrug resistance and is associated with poor prognosis. Therefore, it is important to identify the mechanisms by which MM cells develop multidrug resistance. Our previous study showed that multidrug resistance is correlated with overexpression of multidrug resistance protein 1 (MDR1) and Survivin, and downregulation of Bim expression in melphalan-resistant RPMI8226/L-PAM cells; however, the underlying mechanism of multidrug resistance remains unclear. In the present study, we investigated the mechanism of multidrug resistance in melphalan-resistant cells. We found that RPMI8226/L-PAM and ARH-77/L-PAM cells showed increased phosphorylation of extracellular signal-regulated protein kinase 1/2 (ERK1/2) and Akt, and nuclear localization of nuclear factor κB (NF-κB). The combination of ERK1/2, Akt, and NF-κB inhibitors with melphalan reversed melphalan resistance via suppression of Survivin expression and enhanced Bim expression in melphalan-resistant cells. In addition, RPMI8226/L-PAM and ARH-77/L-PAM cells overexpressed hypoxia-inducible factor 1α (HIF-1α) via activation of ERK1/2, Akt, and NF-κB. Moreover, suppression of HIF-1α by echinomycin or HIF-1α siRNA resensitized RPMI8226/L-PAM cells to melphalan through downregulation of Survivin expression and upregulation of Bim expression. These results indicate that enhanced Survivin expression and decreased Bim expression by HIF-1α via activation of ERK1/2, Akt, and NF-κB play a critical role in melphalan resistance. Our findings suggest that HIF-1α, ERK1/2, Akt, and NF-κB inhibitors are potentially useful as anti-MDR agents for the treatment of melphalan-resistant MM.
  • 婦人科悪性腫瘍におけるパクリタキセル・カルボプラチン療法での後発品製剤安全性の検討(An Analysis of Generic Drug Safety in Paclitaxel and Carboplatin Chemotherapy for Gynecologic Malignancies), 藤本 伸一郎, 柳江 正嗣, 淺野 肇, 武田 朋也, 椿 正寛, 藤原 季美子, 月岡 康行, 松野 純男, 森嶋 祥之, 西田 升三, ジェネリック研究, ジェネリック研究, 12(2), 74 - 79, Dec. 2018
    Summary:わが国では後発医薬品への切り替えが推進されているが、切り替え時の問題点として、臨床試験における同等性の報告が義務付けられていないことが挙げられる。安全域が狭いがん化学療法の分野において、切り替えによる有害事象発現の差異が治療に影響する可能性は高く、実臨床での薬剤評価が必要と考える。しかし、後発医薬品への切り替えに際し、製剤毎での比較報告はあるものの、後発医薬品を併用したレジメンとしての報告は少なく、十分なエビデンスが少ないなかで治療が行われているのが現状である。本検討では婦人科悪性腫瘍における重要なレジメンである、パクリタキセル・カルボプラチン療法における切り替えによる安全性の差異を確認した。結果として、血液毒性、筋肉痛及び関節痛については、切り替え後も同等である可能性が示唆された。他の非血液毒性については、切り替えによりオッズ比は低下傾向であったが、同等性は保証されなかった。また、治療中に減量を必要とした症例数に有意な差は認められず、切り替え後も安全性が保たれていることが示唆された。今後も症例数を重ね、より科学的根拠のある報告をする必要がある。(著者抄録)
  • The MIP-1α autocrine loop contributes to decreased sensitivity to anticancer drugs., Masanobu Tsubaki, Tomoya Takeda, Yoshika Tomonari, Kenji Mashimo, Yu-Ichi Koumoto, Sachi Hoshida, Tatsuki Itoh, Motohiro Imano, Takao Satou, Katsuhiko Sakaguchi, Shozo Nishida, Journal of cellular physiology, Journal of cellular physiology, 233(5), 4258 - 4271, May 2018 , Refereed
    Summary:Several autocrine soluble factors, including macrophage inflammatory protein-1α (MIP-1α), tumor necrosis factor-α, and hepatocyte growth factor, promote cell survival and growth in multiple myeloma (MM) cells. We hypothesized that inhibition of the MIP-1α autocrine loop may enhance the cytotoxic effect of anticancer drugs in MM cell lines. In the present study, an MIP-1α neutralizing antibody suppressed cell proliferation and enhanced the cytotoxic effect of melphalan or bortezomib on MM cells. In addition, melphalan resistance cells (RPMI8226/L-PAM and HS-sultan/L-PAM cells) secreted MIP-1α and neutralizing antibody of MIP-1α partially overcame melphalan resistance. Moreover, combination treatment with MIP-1α neutralizing antibody and melphalan or bortezomib inhibited extracellular signal regulated kinase 1/2 (ERK1/2), Akt, and mammalian target of rapamycin (mTOR) activation, Bcl-2, Bcl-xL, and Survivin expression, and upregulated the expression of Bim and cleaved Poly (ADP-ribose) polymerase (PARP). Treatment of IM9 cells with MIP-1α siRNA suppressed the activation of ERK1/2, Akt, and mTOR, and enhanced the cytotoxic effect of melphalan and bortezomib. These results indicate that MIP-1α neutralizing antibodies or MIP-1α siRNA enhance the cytotoxic effect of melphalan and bortezomib by suppressing the chemokine receptor/ERK and chemokine receptor/Akt/mTOR pathways. The inhibition of MIP-1α may thus provide a new therapeutic approach to control tumor progression and bone destruction in patients with MM.
  • Pioglitazone inhibits cancer cell growth through STAT3 inhibition and enhanced AIF expression via a PPARγ-independent pathway., Masanobu Tsubaki, Tomoya Takeda, Yoshika Tomonari, Keishi Kawashima, Tatsuki Itoh, Motohiro Imano, Takao Satou, Shozo Nishida, Journal of cellular physiology, Journal of cellular physiology, 233(4), 3638 - 3647, Apr. 2018 , Refereed
    Summary:Pioglitazone is an anti-diabetic agent that belongs to the thiazolidinedione class, which target peroxisome proliferator-activated receptor γ (PPARγ), a transcription factor in the nuclear receptor family. Different cancer cells expressing high levels of PPARγ and PPARγ ligands induce cell cycle arrest, cell differentiation, and apoptosis. However, the mechanisms underlying these processes remain unknown. Here, we investigated the mechanism underlying pioglitazone-induced apoptosis in human cancer cells. We showed that at similar concentrations, pioglitazone induced death in cancer cells expressing high or low levels of PPARγ. Combined treatment of pioglitazone and GW9662, a PPARγ antagonist, did not rescue this cell death phenotype. Z-VAD-fmk, a pan-caspase inhibitor, did not reverse pioglitazone-induced apoptosis in cancer cells expressing PPARγ at high or low levels. Pioglitazone suppressed the activation of signal transducers and activator of transcription 3 (STAT3) and Survivin expression, and enhanced the apoptosis-inducing factor (AIF) levels in these cells. Furthermore, pioglitazone enhanced the cytotoxic effect of cisplatin and oxaliplatin by suppressing Survivin and increasing AIF expression. These results indicated that pioglitazone induced apoptosis via a PPARγ-independent pathway, thus describing pioglitazone as a potential therapeutic agent for controlling the progression of different cancers.
  • Comparison of the perception between pharmacy students and practicing pharmacists in the acquisition of physical assessment skills, Toru Otori, Tomomi Inoue, Koichi Hosomi, William Figoni, Manabu Kitakoji, Hiroko Hachiken, Hiroyuki Nakagawa, Keiko Takashima, Hisami Kondo, Tsugumi Takada, Kenji Matsuyama, Shozo Nishida, Japn. J. Soc. Pharm, Japn. J. Soc. Pharm, 37(2), 127 - 133, 2018 , Refereed
  • Mangiferin enhances the sensitivity of human multiple myeloma cells to anticancer drugs through suppression of the nuclear factor κB pathway., Tomoya Takeda, Masanobu Tsubaki, Toshiki Kino, Ayako Kawamura, Shota Isoyama, Tatsuki Itoh, Motohiro Imano, Genzoh Tanabe, Osamu Muraoka, Hideaki Matsuda, Takao Satou, Shozo Nishida, International journal of oncology, International journal of oncology, 48(6), 2704 - 12, Jun. 2016 , Refereed
    Summary:Multiple myeloma (MM) is still an incurable hematological malignancy with a 5-year survival rate of ~35%, despite the use of various treatment options. The nuclear factor κB (NF-κB) pathway plays a crucial role in the pathogenesis of MM. Thus, inhibition of the NF-κB pathway is a potential target for the treatment of MM. In a previous study, we showed that mangiferin suppressed the nuclear translocation of NF-κB. However, the treatment of MM involves a combination of two or three drugs. In this study, we examined the effect of the combination of mangiferin and conventional anticancer drugs in an MM cell line. We showed that the combination of mangiferin and an anticancer drug decreased the viability of MM cell lines in comparison with each drug used separately. The decrease in the combination of mangiferin and an anticancer drug induced cell viability was attributed to increase the expression of p53 and Noxa and decreases the expression of XIAP, survivin, and Bcl-xL proteins via inhibition of NF-κB pathway. In addition, the combination treatment caused the induction of apoptosis, activation of caspase-3 and the accumulation of the cells in the sub-G1 phase of the cell cycle. Our findings suggest that the combination of mangiferin and an anticancer drug could be used as a new regime for the treatment of MM.
  • Statins inhibited the MIP-1α expression via inhibition of Ras/ERK and Ras/Akt pathways in myeloma cells., Masanobu Tsubaki, Kenji Mashimo, Tomoya Takeda, Toshiki Kino, Arisa Fujita, Tatsuki Itoh, Motohiro Imano, Katsuhiko Sakaguchi, Takao Satou, Shozo Nishida, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 78, 23 - 29, Mar. 2016 , Refereed
    Summary:Macrophage inflammatory protein-1alpha (MIP-1α) is detected at high concentrations in patients with multiple myeloma. It is thought to play an important role in the etiology of multiple myeloma and osteolysis. Thus, inhibiting MIP-1α expression may be useful in developing therapeutic treatments for multiple myeloma-induced osteolysis. In this study, we investigated the potential of statins to inhibit mRNA expression and secretion of MIP-1α in mouse myeloma cells (MOPC-31C). We found that statins inhibited the lipopolysaccharide (LPS)-induced MIP-1α mRNA expression and protein secretion in MOPC-31C cells. This inhibition was reversed when farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP), intermediates of the mevalonate pathway, were combined with statins. Furthermore, statins reduced the GTP form of Ras, a phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2), and phosphorylated Akt. Our results indicate that statins inhibit biosynthesis of FPP and GGPP and thereby down regulate signal transduction of Ras/ERK and Ras/Akt pathways. The net effect suppresses LPS-induced MIP-1α mRNA expression and protein secretion in MOPC-31C cells. Thus, statins hold great promise for developing effective therapies against myeloma-induced osteolysis.
  • Overexpression of survivin via activation of ERK1/2, Akt, and NF-κB plays a central role in vincristine resistance in multiple myeloma cells., Masanobu Tsubaki, Tomoya Takeda, Naoki Ogawa, Kotaro Sakamoto, Hirotaka Shimaoka, Arisa Fujita, Tatsuki Itoh, Motohiro Imano, Toshihiko Ishizaka, Takao Satou, Shozo Nishida, Leukemia research, Leukemia research, 39(4), 445 - 52, Apr. 2015 , Refereed
    Summary:The acquisition of anti-cancer drug resistance is a major limitation of chemotherapy for multiple myeloma (MM) and it is thus important to identify the mechanisms by which MM cells develop such drug resistance. In a previous study, we showed that multidrug resistance (MDR) involves the overexpression of MDR1 and survivin in vincristine-resistant RPMI8226/VCR cells. However, the underlying mechanism of MDR remains unclear. In this study, we investigated the mechanism of MDR in RPMI8226/VCR cells, and found that RPMI8226/VCR cells exhibit increased levels of activated ERK1/2, Akt, and NF-κB, while the levels of activated mTOR, p38MAPK, and JNK do not differ between RPMI8226/VCR cells and their vincristine-susceptible counterparts. In addition, the inhibition of ERK1/2, Akt, or NF-κB by inhibitors reversed the drug-resistance of RPMI8226/VCR cells via the suppression of survivin expression, but did not affect MDR1 expression; RNA silencing of survivin expression completely reversed vincristine resistance, while MDR1 silencing only weakly suppressed vincristine resistance in RPMI8226/VCR cells. These results indicate that enhanced survivin expression via the activation of ERK1/2, Akt, and NF-κB plays a critical role in vincristine resistance in RPMI8226/VCR cells. Our findings suggest that ERK1/2, Akt, and NF-κB inhibitors are potentially useful as anti-MDR agents for the treatment of vincristine-resistant MM.
  • Mangiferin suppresses CIA by suppressing the expression of TNF-α, IL-6, IL-1β, and RANKL through inhibiting the activation of NF-κB and ERK1/2., Masanobu Tsubaki, Tomoya Takeda, Toshiki Kino, Tatsuki Itoh, Motohiro Imano, Genzo Tanabe, Osamu Muraoka, Takao Satou, Shozo Nishida, American journal of translational research, American journal of translational research, 7(8), 1371 - 81, 2015 , Refereed
    Summary:Rheumatoid arthritis is a systemic autoimmune disease characterized by chronic inflammation of synovial joints, ultimately leading to a progressive and irreversible joint destruction. Activation of nuclear factor-kappa B (NF-κB) promotes production of proinflammatory cytokines in various inflammatory diseases including rheumatoid arthritis. Mangiferin, 1,3,6,7-tetrahydroxyxanthone-C2-β-D-glucoside (C-glucosyl xanthone), is a naturally occurring polyphenol. Our previous results showed that mangiferin suppressed NF-κB activation. However, it is unclear, whether mangiferin can prevent rheumatoid arthritis through suppression of NF-κB activation and expression of various cytokines, such as tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6), which play a critical role in the pathogenesis of rheumatoid arthritis. In the present study, we found that mangiferin suppressed the progression and incidence of CIA in DBA1/J mice. In CIA mice, mangiferin inhibited the mRNA expression of cytokine genes in thymus and spleen of CIA mie and led to decreased serum levels of IL-1β, IL-6, TNF-α, and receptor activator NF-κB ligand (RANKL) via inhibition of NF-κB and activation of extracellular signal-regulated kinase 1/2 (ERK1/2). In addition, mangiferin markedly inhibited not only developing but also clinically evident CIA. These findings suggest that mangiferin has potential clinical applications for the treatment of rheumatoid arthritis.
  • Bisphosphonates and statins inhibit expression and secretion of MIP-1α via suppression of Ras/MEK/ERK/AML-1A and Ras/PI3K/Akt/AML-1A pathways., Masanobu Tsubaki, Tomoya Takeda, Kotaro Sakamoto, Hirotaka Shimaoka, Arisa Fujita, Tatsuki Itoh, Motohiro Imano, Kenji Mashimo, Daiichiro Fujiwara, Katsuhiko Sakaguchi, Takao Satou, Shozo Nishida, American journal of cancer research, American journal of cancer research, 5(1), 168 - 79, 2015 , Refereed
    Summary:Osteolytic bone disease in multiple myeloma (MM) is associated with upregulated osteoclast activity. Macrophage inflammatory protein-1α (MIP-1α) is crucially involved in the development of osteolytic bone lesions in MM. We previously reported that minodronate inhibited lipopolysaccharide-induced MIP-1α secretion in mouse myeloma cells. However, it remains unknown whether bisphosphonates and statins inhibit MIP-1α secretion by human MM cells. In present study, we investigated whether bisphosphonates and statins had any inhibitory effect on MIP-1α secretion by human myeloma cells and the mechanism underlying this effect. In this study, we found that bisphosphonates and statins inhibited MIP-1α mRNA and MIP-1α secretion and suppressed extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt phosphorylation by inhibiting Ras prenylation. Moreover, bisphosphonates and statins suppressed the expression of acute myeloid leukemia-1A (AML-1A) mRNA, a MIP-1α transcription factor. These results indicate that bisphosphonates and statins suppress the Ras/mitogen-activated protein kinase kinase/ERK/AML-1A and Ras/phosphatidylinositol-3 kinase/Akt/AML-1A pathways, thereby inhibiting MIP-1α secretion by MM cells. Therefore, use of MIP-1α expression inhibitors such as bisphosphonates and statins may provide a new therapeutic approach to inhibiting tumour progression and bone destruction in MM patients.
  • Dimethyl fumarate induces apoptosis of hematopoietic tumor cells via inhibition of NF-κB nuclear translocation and down-regulation of Bcl-xL and XIAP., Masanobu Tsubaki, Naoki Ogawa, Tomoya Takeda, Kotaro Sakamoto, Hirotaka Shimaoka, Arisa Fujita, Tatsuki Itoh, Motohiro Imano, Takao Satou, Shozo Nishida, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 68(8), 999 - 1005, Oct. 2014 , Refereed
    Summary:Dimethyl fumarate (DMF) is a fumaric acid ester that is used to treat psoriasis and multiple sclerosis. Recently, DMF was found to exhibit anti-tumor effects. However, the molecular mechanisms underlying these effects have not been elucidated. In this study, we investigated the mechanism of DMF-induced apoptosis in different human hematopoietic tumor cell lines. We found that DMF induced apoptosis in different human hematopoietic tumor cell lines but it did not affect the normal human B lymphocyte cell line RPMI 1788. We also observed a concurrent increase in caspase-3 activity and in the number of Annexin-V-positive cells. Furthermore, an examination of the survival signals, which are activated by apoptotic stimuli, revealed that DMF significantly inhibited nuclear factor-κB (NF-κB) p65 nuclear translocation. In addition, DMF suppressed B-cell lymphoma extra-large (Bcl-xL) and X-linked inhibitor of apoptosis (XIAP) expression whereas Bcl-2, survivin, Bcl-2-associated X protein (Bax), and Bim levels did not change. These results indicated that DMF induced apoptosis by suppressing NF-κB activation, and Bcl-xL and XIAP expression. These findings suggested that DMF might have potential as an anticancer agent that could be used in combination therapy with other anticancer drugs for the treatment of human hematopoietic tumors.
  • Activation of NF-κB by the RANKL/RANK system up-regulates snail and twist expressions and induces epithelial-to-mesenchymal transition in mammary tumor cell lines., Masanobu Tsubaki, Makiko Komai, Shin-Ichiro Fujimoto, Tatsuki Itoh, Motohiro Imano, Kotaro Sakamoto, Hirotaka Shimaoka, Tomoya Takeda, Naoki Ogawa, Kenji Mashimo, Daiichiro Fujiwara, Junji Mukai, Katsuhiko Sakaguchi, Takao Satou, Shozo Nishida, Journal of experimental & clinical cancer research : CR, Journal of experimental & clinical cancer research : CR, 32(1), 62 - 62, Sep. 05 2013 , Refereed
    Summary:BACKGROUND: Increased motility and invasiveness of cancer cells are reminiscent of the epithelial-mesenchymal transition (EMT), which occurs during cancer progression and metastasis. Recent studies have indicated the expression of receptor activator of nuclear factor-κB (RANK) in various solid tumors, including breast cancer. Although activation of the RANK ligand (RANKL)/RANK system promotes cell migration, metastasis, and anchorage-independent growth of tumor-initiating cells, it remains to be investigated if RANKL induces EMT in breast cancer cells. In this study, we investigated whether RANKL induces EMT in normal breast mammary epithelial cells and breast cancer cells, and the mechanism underlying such induction. METHODS: Expression levels of vimentin, N-cadherin, E-cadherin, Snail, Slug, and Twist were examined by real-time polymerase chain reaction. Cell migration and invasion were assessed using Boyden chamber and invasion assays, respectively. The effects of RANKL on signal transduction molecules were determined by western blot analyses. RESULTS: We found that stimulation by RANKL altered the cell morphology to the mesenchymal phenotype in normal breast epithelial and breast cancer cells. In addition, RANKL increased the expression levels of vimentin, N-cadherin, Snail, and Twist and decreased the expression of E-cadherin. We also found that RANKL activated nuclear factor-κB (NF-κB), but not extracellular signal-regulated kinase 1/2, Akt, mammalian target of rapamycin, c-Jun N-terminal kinase, and signal transducer and activator of transcription 3. Moreover, dimethyl fumarate, a NF-κB inhibitor, inhibited RANKL-induced EMT, cell migration, and invasion, and upregulated the expressions of Snail, Twist, vimentin, and N-cadherin. CONCLUSIONS: The results indicate that RANKL induces EMT by activating the NF-κB pathway and enhancing Snail and Twist expression. These findings suggest that the RANKL/RANK system promotes tumor cell migration, invasion, and metastasis via the induction of EMT.
  • Activation of NF-κB by the RANKL/RANK system up-regulates snail and twist expressions and induces epithelial-to-mesenchymal transition in mammary tumor cell lines., Tsubaki M, Komai M, Fujimoto S, Itoh T, Imano M, Sakamoto K, Shimaoka H, Takeda T, Ogawa N, Mashimo K, Fujiwara D, Mukai J, Sakaguchi K, Satou T, Nishida S, Journal of experimental & clinical cancer research : CR, Journal of experimental & clinical cancer research : CR, 32, 62, Sep. 2013 , Refereed
  • Use of a Vial Form of Fluorouracil to Reduce Occupational Contamination in the Hospital Work Environment and Increase the Efficiency of Mixing Operation, Yoshida Jin, Nakano Hiroyuki, Tei Genshin, Nishida Shozo, Koda Shigeki, Kumagai Shinji, Iryo Yakugaku (Japanese Journal of Pharmaceutical Health Care and Sciences), Iryo Yakugaku (Japanese Journal of Pharmaceutical Health Care and Sciences), 38(7), 409 - 413, Jul. 2012
    Summary:The aim of the present study was to compare contamination by 5-FU between a conventional mixing method of a portable disposable infuser pump using an ampule form of 5-FU and a mixing method of an infuser pump using a vial form of 5-FU. The pharmacist handled 5-FU on the stainless steel tray in the biological safety cabinet. Wipe samples of the stainless steel tray and glove samples from the pharmacist were collected. Then the mixing operation was timed by stop-watch.<br>When the pharmacist prepared the infuser pump with ample form of 5-FU, the median 5-FU concentration from wipe samples was 33 μg. When the pharmacist prepared the infuser pump with the vial form of 5-FU, the median 5-FU contamination from wipe samples was lower than the quantification limit. Use of the vial form of 5-FU significantly reduced the surface contamination from wipe samples (<i>p</i>=0.003, Mann-Whitney's <i>U</i>-test). The median mixing operation time was also significantly reduced by using the vial form of 5-FU (347 sec and 169 sec, <i>p</i><0.001, Mann-Whitney's <i>U</i>-test). Based on these results, we concluded that the vial form of 5-FU can reduce contamination in a hospital work environment, and increase the efficiency of mixing operation.
  • Blockade of the Ras/MEK/ERK and Ras/PI3K/Akt pathways by statins reduces the expression of bFGF, HGF, and TGF-β as angiogenic factors in mouse osteosarcoma., Masanobu Tsubaki, Yuzuru Yamazoe, Masashi Yanae, Takao Satou, Tatsuki Itoh, Junichi Kaneko, Yasuhiro Kidera, Kenzo Moriyama, Shozo Nishida, Cytokine, Cytokine, 54(1), 100 - 7, Apr. 2011 , Refereed
    Summary:The tumor microenvironment plays a critical role in modulating malignant behavior and can dramatically influence cancer treatment strategies. We investigated whether statins inhibit the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and transforming growth factor-β (TGF-β) mRNA in the mouse osteosarcoma cell line LM8. We found that statins significantly inhibited mRNA expressions of bFGF, HGF, and TGF-β, and bFGF, HGF, and TGF-β secretions at concentrations that did not have antiproliferative effects on LM8 cells, but had no effect on the mRNA expression and secretion of VEGF. The inhibition of bFGF, HGF, and TGF-β mRNA expression, and bFGF, HGF, TGF-β secretions was reversed when geranylgeranyl pyrophosphate (GGPP), an intermediate in the mevalonate pathway, was used in combination with statins. Furthermore, statins reduced the membrane localization of K-Ras, phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2), and phosphorylated Akt. Our research indicates that statins inhibit GGPP biosynthesis in the mevalonate pathway, and then inhibit signal transduction in the Ras/ERK and Ras/Akt pathways, thereby inhibiting bFGF, HGF, TGF-β expression in LM8 cells. These results suggest that statins are potentially useful as anti-angiogenic agents for the treatment of osteosarcoma.
  • Proposal of Checklist for Safe Handling of Antineoplastic Drugs, Yoshida Jin, Koda Shigeki, Yoshida Toshiaki, Nishida Syozo, Kumagai Shinji, Iryo Yakugaku (Japanese Journal of Pharmaceutical Health Care and Sciences), Iryo Yakugaku (Japanese Journal of Pharmaceutical Health Care and Sciences), 37(3), 145 - 155, Mar. 2011
    Summary:This study was conducted to develop a tool for evaluating the work environment for the preparation of antineoplasticdrugs and identify issues in safety assessment.We prepared a checklist for the safe handling of antineoplastic drugs,whichincluded the following items : A) safety equipment and its maintenance,B) training and training documentation,C) devicesfor ensuring safety,D) personal protective equipment and E) emergency care.Further,we examined relationships betweenattainment levels for checklist items and occupational contamination levels for cyclophosphamide (CPA),fluorouracil (5FU),gemcitabine (GEM) and platinum-containing drugs (Pt) based on wipe samples from equipment in the preparationroom (wipe test).Next,pharmacists in a particular hospital were asked to improve their work environment using the checklist.To evaluate their improvements,we collected wipe samples and one-day urine samples from them,before and afterthey took improvement measures.The results showed that an increase in safety attainment scores was reflected in a decrease in CPA,GEM and Pt contaminationlevels in the wipe test,as well as one in urinary levels of CPA.However,the 5FU level in the wipe test increased,probably due to a 6-fold increase in the amount of 5FU prepared,compared to before the improvement measures weretaken.From these findings,we concluded that the checklist would be useful for improving the hospital work environment.
  • Mangiferin induces apoptosis by suppressing Bcl-xL and XIAP expressions and nuclear entry of NF-κB in HL-60 cells., Kaori Shoji, Masanobu Tsubaki, Yuzuru Yamazoe, Takao Satou, Tatsuki Itoh, Yasuhiro Kidera, Yoshihiro Tanimori, Masashi Yanae, Hideaki Matsuda, Atsushi Taga, Haruyuki Nakamura, Shozo Nishida, Archives of pharmacal research, Archives of pharmacal research, 34(3), 469 - 75, Mar. 2011 , Refereed
    Summary:Mangiferin, 1,3,6,7-tetrahydroxyxanthone-C2-β-D: -glucoside (C-glucosylxanthone), is a xanthone derivative that is widely distributed in higher plants. Recently, mangiferin was found to exhibit potential antitumor effects. However, the molecular mechanisms of this effect have not been elucidated. In the present study, we attempt to clarify the mechanism of mangiferin-induced apoptosis in the human acute myeloid leukemia cell line HL-60; mangiferin was found to induce apoptosis. We also observed a concurrent increase in caspase-3 activity and DNA fragmentation. Furthermore, on examining the survival signals expressed during apoptotic induction, we observed that mangiferin caused a remarkable decrease in the nuclear entry of NF-κB p65. However, there were no changes in the expression of other survival signals, such as extracellular signal-regulated kinase 1/2, protein kinase B, and p38 mitogenactivated protein kinase. In addition, mangiferin suppressed the expressions of Bcl-xL and XIAP; however, we did not note any changes in the levels of Bcl-2, Bax, and Bim. These results indicate that mangiferin induces apoptosis by suppressing NF-κB activation and expressions of Bcl-xL and XAIP. These findings suggest that mangiferin may be useful as an anticancer agent and can be used in combination therapy with other anticancer drugs for the treatment of acute myeloid leukemia.
  • Expression and Cerebral function of amyloid precursor protein after rat traumatic brain injury., 31 - 52, 2011 , Refereed
  • Macrophage inflammatory protein-1α induces osteoclast formation by activation of the MEK/ERK/c-Fos pathway and inhibition of the p38MAPK/IRF-3/IFN-β pathway., Masanobu Tsubaki, Chisato Kato, Ai Isono, Junichi Kaneko, Misako Isozaki, Takao Satou, Tatsuki Itoh, Yasuhiro Kidera, Yoshihiro Tanimori, Masashi Yanae, Shozo Nishida, Journal of cellular biochemistry, Journal of cellular biochemistry, 111(6), 1661 - 72, Dec. 15 2010 , Refereed
    Summary:Multiple myeloma (MM) is a bone disease that affects many individuals. It was recently reported that macrophage inflammatory protein (MIP)-1α is constitutively secreted by MM cells. MIP-1α causes bone destruction through the formation of osteoclasts (OCs). However, the molecular mechanism underlying MIP-1α-induced OC formation is not well understood. In the present study, we attempted to clarify the mechanism whereby MIP-1α induces OC formation in a mouse macrophage-like cell line comprising C7 cells. We found that MIP-1α augmented OC formation in a concentration-dependent manner; moreover, it inhibited IFN-β and ISGF3γ mRNA expression, and IFN-β secretion. MIP-1α increased the expressions of phosphorylated ERK1/2 and c-Fos and decreased those of phosphorylated p38MAPK and IRF-3. We found that the MEK1/2 inhibitor U0126 inhibited OC formation by suppressing the MEK/ERK/c-Fos pathway. SB203580 induced OC formation by upregulating c-fos mRNA expression, and SB203580 was found to inhibit IFN-β and IRF-3 mRNA expressions. The results indicate that MIP-1α induces OC formation by activating and inhibiting the MEK/ERK/c-Fos and p38MAPK/IRF-3 pathways, respectively, and suppressing IFN-β expression. These findings may be useful in the development of an OC inhibitor that targets intracellular signaling factors.
  • The chronological changes for stromal cell-derived factor-1α after rat traumatic brain injury, Neurotrauma Research, Neurotrauma Research, 18, 8 - 12, 2006
  • A case of possible thyroid metastasis from lung carcinoma, 47, 367 - 373, 2002
  • An immunohistochemical study of Fas ligande in head and neck squamous cell carcinoma, 47(5), 367 - 373, Sep. 2001
  • Mn-SOD in human otitis media, 46(2), 96 - 100, Mar. 2000
  • Regional distribution of superoxide dismutase in the brain and myocardium of the stroke-prone spontaneously hypertensive rat., Kimoto S, Nishida S, Funasaka K, Nakano T, Teramoto K, Tomura TT, Clinical and experimental pharmacology & physiology. Supplement, Clinical and experimental pharmacology & physiology. Supplement, 22, S160 - 1, Dec. 1995 , Refereed
  • Dimethyl fumarate suppresses metastasis and growth of melanoma cells by inhibiting the nuclear translocation of NF-κB., Tomoya Takeda, Masanobu Tsubaki, Ryota Asano, Tatsuki Itoh, Motohiro Imano, Takao Satou, Shozo Nishida, Journal of dermatological science, Journal of dermatological science, Jul. 16 2020 , Refereed
    Summary:BACKGROUND: Malignant melanoma is among the deadliest forms of skin cancers, and its incidence has been increasing over the past decades. In malignant melanoma, activation of the nuclear factor kappa B (NF-κB) promotes survival, migration, and invasion of cancer cells. Anti-NF-κB agents for treating metastatic melanoma would be beneficial, but no such drug is approved as either monotherapy or adjuvant therapy. Dimethyl fumarate (DMF) is an approved anti-inflammatory drug already in clinical use for psoriasis and multiple sclerosis. OBJECTIVE: We investigated the anti-tumour effect of DMF treatment in metastatic melanoma in vitro and in vivo. METHODS: The cell viability was assessed via trypan blue exclusion assay. The migration and invasion was analyzed in a Boyden chamber assay. The anti-metastatic effects and anti-tumour activity of DMF was determined in an in-vivo model. The expressions of NF-κB pathway and NF-κB regulatory proteins were detected via western blotting. RESULTS: DMF decreased the cell viability, migration and invasion in vitro. In addition, DMF inhibited spontaneous metastasis and tumour growth. Mechanistically, DMF prevented the nuclear translocation of NF-κB, whereas no changes were observed in the phosphorylation levels of inhibitor of kappa B (IκB). In addition, DMF inhibited the expression of matrix metalloproteinases (MMPs) and very late antigens (VLAs). Furthermore, DMF treatment decreased the expression of Survivin and Bcl-extra large (Bcl-XL) proteins. CONCLUSION: Our results suggest that DMF as a novel inhibitor of NF-κB may be a potential therapeutic agent for metastatic melanoma.
  • [Three Cases of Augmented Chemotherapy-Induced Peripheral Neuropathy after Changing from mFOLFOX6 to FOLFIRI Therapy in Patients with Colorectal Cancer]., Keiji Mashimo, Daichiro Fujiwara, Tadafumi Hoshida, Naomi Morimoto, Akihiro Noda, Tomoya Takeda, Masanobu Tsubaki, Shozo Nishida, Katsuhiko Sakaguchi, Gan to kagaku ryoho. Cancer & chemotherapy, Gan to kagaku ryoho. Cancer & chemotherapy, 47(6), 993 - 995, Jun. 2020 , Refereed
    Summary:We had cases in which peripheral neuropathy was augmented after changing from mFOLFOX6, a chemotherapy for colorectal cancer, to FOLFIRI and comparatively examined disease status and trends. There were no shared points with respect to patient characteristics, timing of peripheral neuropathy augmentation, drug dosage, etc. It appeared that the change in chemotherapy itself had an effect on neuropathic symptoms. Regarding the change in chemotherapy, the therapeutic agent was switched from oxaliplatin to irinotecan; the cause was unknown, but some effects of these two drugs were suggested. Future investigation, including the examination of genetic mutations, is necessary.
  • Dasatinib reverses drug resistance by downregulating MDR1 and Survivin in Burkitt lymphoma cells., Mitsuki Tabata, Masanobu Tsubaki, Tomoya Takeda, Keisuke Tateishi, Katsumasa Tsurushima, Motohiro Imano, Takao Satou, Toshihiko Ishizaka, Shozo Nishida, BMC complementary medicine and therapies, BMC complementary medicine and therapies, 20(1), 84 - 84, Mar. 14 2020 , Refereed
    Summary:BACKGROUND: Current chemotherapies for Burkitt lymphoma (BL) have dramatically improved its clinical outcome. However, chemoresistance can lead to chemotherapy failure and very poor prognosis; thus, novel strategies are urgently required for patients with drug-resistant BL. To investigate the mechanisms underlying drug resistance in BL, we established drug-resistant BL cell lines: HS-Sultan/ADM (adriamycin-resistant), HS-Sultan/VCR (vincristine-resistant), HS-Sultan/DEX (dexamethasone-resistant), and HS-Sultan/L-PAM (melphalan-resistant). METHODS: Drug transporter and survival factor expression were investigated the using western blotting and real time polymerase chain reaction. Cell survival was analyzed by trypan blue dye exclusion method. RESULTS: The established cell lines acquired cross-resistance to adriamycin, vincristine, dexamethasone, and melphalan and exhibited 50% inhibitory concentration values 106-, 40-, 81-, and 45-fold higher than the parental cell lines, respectively. We found that protein and mRNA expression of MDR1 and Survivin were higher in drug-resistant BL cells than in the parent cells. Treatment with verapamil, an MDR1 inhibitor, or Survivin siRNA alongside each anti-cancer drug suppressed the proliferation of all drug-resistant BL cells. Src kinase activity was higher in all resistant cell lines than the parental cells; suppressing Src with dasatinib restored drug sensitivity by reducing MDR1 and Survivin expression. CONCLUSIONS: MDR1 and Survivin upregulation are responsible for resistance to conventional drugs and dasatinib can restore drug sensitivity by reducing MDR1 and Survivin expression in drug-resistant BL cells. Src inhibitors could therefore be a novel treatment strategy for patients with drug resistant BL.
  • Inhibition of HSP90 overcomes melphalan resistance through downregulation of Src in multiple myeloma cells., Mitsuki Tabata, Masanobu Tsubaki, Tomoya Takeda, Keisuke Tateishi, Saho Maekawa, Katsumasa Tsurushima, Motohiro Imano, Takao Satou, Toshihiko Ishizaka, Shozo Nishida, Clinical and experimental medicine, Clinical and experimental medicine, 20(1), 63 - 71, Feb. 2020 , Refereed
    Summary:Multiple myeloma (MM) is the second most common hematologic malignancy. In spite of the development of new therapeutic agents, MM remains incurable due to multidrug resistance (MDR) and the 5-year survival rate is approximately 50%. Thus, further study is needed to investigate the mechanism of MDR and improve MM prognosis. Heat shock protein 90 (HSP90) is a molecular chaperone that is responsible for the stability of a number of client proteins, most of which are involved in tumor progression. Therefore, HSP90 inhibitors represent potential new therapeutic agents for cancer. Furthermore, inhibition of HSP90 leads to degradation of client proteins, overcoming acquired anti-cancer drug resistance. In this study, we assessed the role of HSP90 in MDR using established melphalan-resistant MM cells. We found that expression of HSP90 was higher in melphalan-resistant MM cells than in parent cells and that HSP90 inhibitors KW-2478 and NUV-AUY922 restored drug sensitivity to the level observed in parent cells. Activation of the unfolded protein response is a hallmark of MM, and expression of endoplasmic reticulum stress signaling molecules is reduced in melphalan-resistant cells; however, KW-2478 did not affect endoplasmic reticulum stress signaling. We demonstrated that treatment with KW-2478 decreased expression of Src, a client of HSP90, and suppressed the activity of ERK, Akt, and NF-κB. Our findings indicate that inhibition of HSP90 results in suppression of Src and its downstream effectors, including ERK, Akt, and NF-κB, and therefore that HSP90 inhibitors could be useful for treatment of MDR MM.
  • Overactivation of Akt Contributes to MEK Inhibitor Primary and Acquired Resistance in Colorectal Cancer Cells., Masanobu Tsubaki, Tomoya Takeda, Masaki Noguchi, Minami Jinushi, Shiori Seki, Yuusuke Morii, Kazunori Shimomura, Motohiro Imano, Takao Satou, Shozo Nishida, Cancers, Cancers, 11(12), Nov. 25 2019 , Refereed
    Summary:RAS and BRAF-mutated colorectal cancers are associated with resistance to chemotherapy and poor prognosis, highlighting the need for new therapeutic strategies. Although these cancers sometimes respond to mitogen activated protein kinase kinase (MEK) inhibitor treatment, they often acquire resistance via mechanisms, which are poorly understood. Here, we investigated the mechanism of MEK inhibitor resistance in primary- and acquired-resistant cells. Cell viability was examined using the trypan blue dye exclusion assay. Protein expression was analyzed by western blotting. Somatic mutations in colorectal cancer cells were investigated using the polymerase chain reaction array. PD0325901 and trametinib induced cell death in LoVo and Colo-205 cells but not in DLD-1 and HT-29 cells, which have a PIK3CA mutation constitutively activating Akt and NF-κB. Treatment with PD0325901 and trametinib suppressed ERK1/2 activation in all four cell lines but only induced Akt and NF-κB activation in DLD-1 and HT-29 cells. Inhibition of Akt but not NF-κB, overcame MEK inhibitor resistance in DLD-1 and HT-29 cells. Acquired-resistant LoVo/PR, Colo-205/PR and LoVo/TR cells have constitutively active Akt due to a M1043V mutation in the kinase activation loop of PIK3CA and Akt inhibitor resensitized these cells to MEK inhibitor. These results demonstrate that the overactivation of Akt plays a critical role in MEK inhibitor primary and acquired resistance and implicate combined Akt/MEK inhibition as a potentially useful treatment for RAS/BRAF-mutated colorectal cancer.
  • Combination therapy with dacarbazine and statins improved the survival rate in mice with metastatic melanoma., Masanobu Tsubaki, Tomoya Takeda, Naoya Obata, Keishi Kawashima, Mitsuki Tabata, Motohiro Imano, Takao Satou, Shozo Nishida, Journal of cellular physiology, Journal of cellular physiology, 234(10), 17975 - 17989, Aug. 2019 , Refereed
    Summary:Malignant melanoma is a highly aggressive skin cancer, and the overall median survival in patients with metastatic melanoma is only 6-9 months. Although molecular targeted therapies have recently been developed and have improved the overall survival, melanoma patients may show no response and acquisition of resistance to these drugs. Thus, other molecular approaches are essential for the treatment of metastatic melanoma. In the present study, we investigated the effect of cotreatment with dacarbazine and statins on tumor growth, metastasis, and survival rate in mice with metastatic melanomas. We found that cotreatment with dacarbazine and statins significantly inhibited tumor growth and metastasis via suppression of the RhoA/RhoC/LIM domain kinase/serum response factor/c-Fos pathway and enhanced p53, p21, p27, cleaved caspase-3, and cleaved poly(ADP-ribose) polymerase 1 expression in vivo. Moreover, the cotreatment significantly improved the survival rate in metastasis-bearing mice. Importantly, treatment with dacarbazine plus 100 mg/kg simvastatin or fluvastatin prevented metastasis-associated death in 4/20 mice that received dacarbazine + simvastatin and in 8/20 mice that received dacarbazine + fluvastatin (survival rates, 20% and 40%, respectively). These results suggested that cotreatment with dacarbazine and statins may thus serve as a new therapeutic approach to control tumor growth and metastasis in melanoma patients.
  • RANKL-induced c-Src activation contributes to conventional anti-cancer drug resistance and dasatinib overcomes this resistance in RANK-expressing multiple myeloma cells., Keiji Mashimo, Masanobu Tsubaki, Tomoya Takeda, Ryota Asano, Minami Jinushi, Motohiro Imano, Takao Satou, Katsuhiko Sakaguchi, Shozo Nishida, Clinical and experimental medicine, Clinical and experimental medicine, 19(1), 133 - 141, Feb. 2019 , Refereed
    Summary:The survival and growth of multiple myeloma (MM) cells are facilitated by cell-cell interactions with bone marrow stromal cells and the bone marrow microenvironment. These interactions induce de novo drug resistance known as cell adhesion-mediated drug resistance. Our previous results recently revealed that the receptor activator of NF-κB (RANK) ligand (RANKL), which is expressed by bone marrow stromal cells, contributes to anti-cancer drug resistance through the activation of various signaling molecules and suppression of Bim expression in RANK-expressing MM cells. However, the detailed mechanisms underlying RANKL-induced drug resistance remain uncharacterized. In the present study, we investigated the mechanism of RANKL-induced drug resistance in RANK-expressing MM cell lines. We found treatment of MM cells with RANKL-induced c-Src phosphorylation and activation of the downstream signaling molecules Akt, mTOR, STAT3, JNK, and NF-κB. In addition, treatment with dasatinib, a c-Src inhibitor, overcame RANKL- and bone marrow stromal cell-induced drug resistance to adriamycin, vincristine, dexamethasone, and melphalan by suppressing c-Src, Akt, mTOR, STAT3, JNK, and NF-κB activation and enhancing expression of Bim. Overall, RANKL- and bone marrow stromal cell-induced drug resistance correlated with the activation of c-Src signaling pathways, which caused a decrease in Bim expression. Dasatinib treatment of RANK-expressing MM cells re-sensitized them to anti-cancer drugs. Therefore, inhibition of c-Src may be a new therapeutic approach for overcoming RANKL-induced drug resistance in patients with MM.
  • Phase II trial of neoadjuvant chemotherapy with intraperitoneal paclitaxel, S-1, and intravenous cisplatin and paclitaxel for stage IIIA or IIIB gastric cancer., Masayuki Shinkai, Motohiro Imano, Yasutaka Chiba, Mitsuru Iwama, Osamu Shiraisi, Atsushi Yasuda, Masanobu Tsubaki, Shozo Nishida, Yutaka Kimura, Takushi Yasuda, Journal of surgical oncology, Journal of surgical oncology, 119(1), 56 - 63, Jan. 2019 , Refereed
    Summary:BACKGROUND: We carried out a phase II trial to evaluate the feasibility and efficacy of neoadjuvant chemotherapy comprising a single intraperitoneal administration of paclitaxel, followed by intravenous administrations of paclitaxel and cisplatin with S-1 for clinical stage III gastric cancer. METHODS: Patients with potentially resectable gastric cancer were eligible. A laparoscopic survey was performed to confirm CY0 and P0. Intraperitoneal paclitaxel (60 mg/m 2 ) was administered, followed by systemic chemotherapy. Surgery was performed after two cycles of chemotherapy. The primary endpoint was the response rate of chemotherapy. Secondary endpoints were adverse events, pathological response rate, and overall survival rate. RESULTS: Twenty patients were enrolled. Planned cycles were completed in all patients. Grade 3/4 leukopenia and grade 3/4 neutropenia were observed in four (20%) and seven (35%) patients, respectively. The overall response rate was 70% (partial response: 14, stable disease: 5, progressive disease: 1). All patients underwent R0 gastrectomy with D2 lymph-node dissection, with no surgery-related deaths. The pathological response rate was 65% (13 of 20). The 3- and 5-year overall survival rates were 90.0% and 77.1%, respectively. CONCLUSIONS: Neoadjuvant chemotherapy including intraperitoneal paclitaxel followed by sequential intravenous paclitaxel and cisplatin with S-1 for resectable advanced gastric cancer is feasible and effective.
  • Intraperitoneal Administration of Paclitaxel Followed by Paclitaxel, Cisplatin, and S-1 Chemotherapy for Cytology-positive Gastric Cancer: A Feasibility Study., Masayuki Shinkai, Motohiro Imano, Yasutaka Chiba, Yoko Hiraki, Hiroaki Kato, Mitsuru Iwama, Osamu Shiraisi, Atsushi Yasuda, Masahiro Tsubaki, Shozo Nishida, Yutaka Kimura, Takushi Yasuda, Anticancer research, Anticancer research, 38(10), 5969 - 5974, Oct. 2018 , Refereed
    Summary:BACKGROUND/AIM: A preliminary study evaluating the feasibility of single intraperitoneal (IP) administration of paclitaxel followed by paclitaxel and cisplatin with S-1 (PCS) systemic chemotherapy for cytology-positive (CY1) gastric cancer. PATIENTS AND METHODS: Staging laparoscopy was performed to confirm CY1 and P0 status. Initially, patients received IP paclitaxel. Beginning 7 days later PCS was given every 3 weeks followed by second-look laparoscopy. RESULTS: Nine patients were enrolled. The toxic effects of IP and systemic chemotherapy were acceptable. After chemotherapy, 8 patients converted from CY1P0 to CY0P0 and 1 patient from CY1P0 to CY1P1. Gastrectomy was performed on 8 patients except for the CY1P1 patient. Four patients were alive without recurrence. The 2-year overall and progression-free survival rates were 76% and 65%, respectively. CONCLUSION: Combination chemotherapy with IP paclitaxel and sequential PCS is safe and may be effective for CY1 gastric cancer.
  • Intraperitoneal and Systemic Chemotherapy for Patients with Gastric Cancer with Peritoneal Metastasis: A Phase II Trial., Masayuki Shinkai, Motohiro Imano, Yasutaka Chiba, Yoko Hiraki, Hiroaki Kato, Mitsuru Iwama, Osamu Shiraishi, Atsushi Yasuda, Masahiro Tsubaki, Shozo Nishida, Yutaka Kimura, Takushi Yasuda, Anticancer research, Anticancer research, 38(10), 5975 - 5981, Oct. 2018 , Refereed
    Summary:AIM: To conduct a phase II study of single intraperitoneal (i.p.) administration of paclitaxel followed by paclitaxel, cisplatin, and S-1 (PCS) chemotherapy for patients with gastric cancer with peritoneal metastasis (PM). PATIENTS AND METHODS: Staging laparotomy was performed to confirm PM. Initially, patients received i.p. paclitaxel. Beginning 7 days later, PCS was given every 3 weeks followed by second-look laparoscopy. Primary and secondary endpoints were the overall survival (OS) rate, and response rate and patient safety, respectively. RESULTS: Seventeen patients were enrolled. The overall response rate was 70.5% (12/17). Grade 3/4 toxic effects included neutropenia and leukopenia. After chemotherapy, PM disappearance was confirmed in 11 patients. Gastrectomy was eventually performed in 11 patients. The 1-year OS rate was 82.4% and the median survival time was 23.9 months considering the overall cohort. CONCLUSION: Combination chemotherapy with i.p. paclitaxel and PCS is well tolerated and effective in patients with gastric cancer with PM.
  • Tamoxifen suppresses paclitaxel-, vincristine-, and bortezomib-induced neuropathy via inhibition of the protein kinase C/extracellular signal-regulated kinase pathway., Masanobu Tsubaki, Tomoya Takeda, Mikihiro Matsumoto, Natsuki Kato, Shota Yasuhara, Yu-Ichi Koumoto, Motohiro Imano, Takao Satou, Shozo Nishida, Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, 40(10), 1010428318808670 - 1010428318808670, Oct. 2018 , Refereed
    Summary:Chemotherapy-induced neuropathy is a highly problematic, dose-limiting effect of potentially curative regimens of cancer chemotherapy. When neuropathic pain is severe, patients often either switch to less-effective chemotherapy agents or choose to discontinue chemotherapy entirely. Conventional chemotherapy drugs used to treat lung and breast cancer, multiple myeloma, and lymphoma include paclitaxel, vincristine, and bortezomib. Approximately 68% of patients receiving these anticancer drugs develop neuropathy within the first month of treatment, and while strategies to prevent chemotherapy-induced neuropathy have been investigated, none have yet been proven as effective. Recent reports suggest that chemotherapy-induced neuropathy is associated with signal transduction molecules, including protein kinase C and mitogen-activated protein kinases. It is currently unclear whether protein kinase C inhibition can prevent chemotherapy-induced neuropathy. In this study, we found that tamoxifen, a protein kinase C inhibitor, suppressed paclitaxel-, vincristine-, and bortezomib-induced cold and mechanical allodynia in mice. In addition, chemotherapy drugs induce neuropathy via the protein kinase C/extracellular signal-regulated kinase pathway in the spinal cord in lumbar segments 4-6 and dorsal root ganglions. In addition, tamoxifen was shown to act synergistically with paclitaxel to inhibit tumor-growth in mice injected with tumor cells. Our results indicated that paclitaxel-, vincristine-, and bortezomib-induced neuropathies were associated with the protein kinase C/extracellular signal-regulated kinase pathway in the lumbar spinal cord and dorsal root ganglions, which suggest that protein kinase C inhibitors may be therapeutically effective for the prevention of chemotherapy-induced neuropathy when administered with standard chemotherapy agents.
  • Bavachin induces the apoptosis of multiple myeloma cell lines by inhibiting the activation of nuclear factor kappa B and signal transducer and activator of transcription 3., Tomoya Takeda, Masanobu Tsubaki, Yoshika Tomonari, Keishi Kawashima, Tatsuki Itoh, Motohiro Imano, Takao Satou, Shozo Nishida, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 100, 486 - 494, Apr. 2018 , Refereed
    Summary:Bavachin is a phytoestrogen purified from natural herbal plants such as Psoralea corylifolia. In this study, we examined the effect of bavachin in multiple myeloma (MM) cell lines. We found that bavachin decreased the viability of MM cell lines, but was not cytotoxic towards normal cells. It inhibited the activation of nuclear factor kappa B (NF-κB) and signal transducer and activator of transcription 3 (STAT3). Furthermore, bavachin increased the expression of p53 and NOXA, and decreased the expression of X-linked inhibitor of apoptosis protein (XIAP), survivin, B cell lymphoma-extra large (Bcl-xL), and Bcl-2. Additionally, bavachin induced apoptosis by the activation of caspase-3 and caspase-9, implicating the involvement of the mitochondrial pathway. Our results suggest that bavachin induces apoptosis through the inhibition of NF-κB and STAT3 activation in MM cell lines. Most importantly, few NF-κB and STAT3 inhibitors with high efficiency, specificity, and safety are currently available for clinical cancer therapy. Hence, bavachin, which targets NF-κB and STAT3, is a potential anticancer agent for the treatment of MM.
  • Rebamipide suppresses 5-fluorouracil-induced cell death via the activation of Akt/mTOR pathway and regulates the expression of Bcl-2 family proteins., Masanobu Tsubaki, Tomoya Takeda, Ryo-Ta Asano, Tomoyuki Matsuda, Shin-Ichiro Fujimoto, Tatsuki Itoh, Motohiro Imano, Takao Satou, Shozo Nishida, Toxicology in vitro : an international journal published in association with BIBRA, Toxicology in vitro : an international journal published in association with BIBRA, 46, 284 - 293, Feb. 2018 , Refereed
    Summary:Oral mucositis is a common adverse effect of chemotherapy that limits the required dose of chemotherapeutic agents. Numerous attempts to mitigate chemotherapy-induced oral mucositis have failed to identify an appropriate treatment. Recently, it has been indicated that rebamipide prevents chemoradiotherapy-induced oral mucositis in patients. However, the details of the underlying mechanism involved in the cytoprotective effect of rebamipide remain obscure. In the present study, we investigated the mechanism behind rebamipide cytoprotective effect in the oral mucosa using primary normal human oral keratinocytes (NHOK cells). We found that rebamipide prevented 5-fluorouracil (5-FU)-induced cell death in NHOK cells. In addition, rebamipide increased the levels of phosphorylated Akt and mTOR, enhanced the Bcl-2 and Bcl-xL expressions, and suppressed the expression of Bax and Bim. This is in contrast to 5-FU-induced suppression of Akt and mTOR activation, Bcl-2 and Bcl-xL expressions, and the enhanced expression of Bax and Bim. These findings suggest that rebamipide can potentially be used for the protection of oral mucosa from chemotherapy-induced mucositis. This is the first study that elucidates the specific molecular pathway for the cytoprotective effect of rebamipide.
  • Trametinib suppresses chemotherapy-induced cold and mechanical allodynia via inhibition of extracellular-regulated protein kinase 1/2 activation., Masanobu Tsubaki, Tomoya Takeda, Mikihiro Matsumoto, Natsuki Kato, Ryo-Ta Asano, Motohiro Imano, Takao Satou, Shozo Nishida, American journal of cancer research, American journal of cancer research, 8(7), 1239 - 1248, 2018 , Refereed
    Summary:Chemotherapy-induced neuropathy is a common, dose-dependent adverse effect of some anti-cancer drugs and leads to discontinuation of chemotherapy and detrimental dose reductions, thereby affecting the quality of life of cancer patients. Currently, no treatment can effectively prevent or treat chemotherapy-induced neuropathy. Therefore, understanding its underlying molecular mechanisms may help to identify novel therapies for treating it. Some disease-induced neuropathy involve the activation of mitogen-activated protein kinases (MAPKs), such as extracellular-regulated protein kinase 1/2 (ERK1/2). In the present study, we investigated whether ERK1/2 inhibition can prevent chemotherapy-induced neuropathy. We found that trametinib, an MEK inhibitor, suppressed oxaliplatin-, paclitaxel-, vincristine-, and bortezomib-induced cold and mechanical allodynia in mice. In addition, treatment with oxaliplatin, paclitaxel, vincristine, or bortezomib enhanced ERK1/2 and c-Jun N-terminal kinase (JNK) phosphorylation in the spinal cord lumbar segments 4-6, and when combined with trametinib, can prevent chemotherapy-induced neuropathy via the suppression of ERK1/2 activation, but does not affect JNK activation. In conclusion, we demonstrated that the disruption of this pathway by MEK inhibitors suppresses oxaliplatin-, paclitaxel-, vincristine-, and bortezomib-induced neuropathy. This suggests that inhibition of the MEK/ERK pathway could prevent chemotherapy-induced neuropathy and MEK inhibitors could be used in combination with anti-tumor drugs during pharmacotherapy.
  • Statins induce apoptosis through inhibition of Ras signaling pathways and enhancement of Bim and p27 expression in human hematopoietic tumor cells., Daichiro Fujiwara, Masanobu Tsubaki, Tomoya Takeda, Yoshika Tomonari, Yu-Ichi Koumoto, Katsuhiko Sakaguchi, Shozo Nishida, Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, 39(10), 1010428317734947 - 1010428317734947, Oct. 2017 , Refereed
    Summary:Recently, statins have been demonstrated to improve cancer-related mortality or prognosis in patients of various cancers. However, the details of the apoptosis-inducing mechanisms remain unknown. This study showed that the induction of apoptosis by statins in hematopoietic tumor cells is mediated by mitochondrial apoptotic signaling pathways, which are activated by the suppression of mevalonate or geranylgeranyl pyrophosphate biosynthesis. In addition, statins decreased the levels of phosphorylated extracellular signal-regulated kinase 1/2 and mammalian target of rapamycin through suppressing Ras prenylation. Furthermore, inhibition of extracellular signal-regulated kinase 1/2 and mammalian target of rapamycin by statins induced Bim expression via inhibition of Bim phosphorylation and ubiquitination and cell-cycle arrest at G1 phase via enhancement of p27 expression. Moreover, combined treatment of U0126, a mitogen-activated protein kinase kinase 1/2 inhibitor, and rapamycin, a mammalian target of rapamycin inhibitor, induced Bim and p27 expressions. The present results suggested that statins induce apoptosis by decreasing the mitochondrial transmembrane potential, increasing the activation of caspase-9 and caspase-3, enhancing Bim expression, and inducing cell-cycle arrest at G1 phase through inhibition of Ras/extracellular signal-regulated kinase and Ras/mammalian target of rapamycin pathways. Therefore, our findings support the use of statins as potential anticancer agents or concomitant drugs of adjuvant therapy.
  • Increased risk of SSEs in bone-only metastatic breast cancer patients treated with zoledronic acid, Masashi Yanae, Shinichiro Fujimoto, Kaori Tane, Maki Tanioka, Kimiko Fujiwara, Masanobu Tsubaki, Yuzuru Yamazoe, Yoshiyuki Morishima, Yasutaka Chiba, Shintaro Takao, Yoshifumi Komoike, Junji Tsurutani, Kazuhiko Nakagawa, Shozo Nishida, Journal of Bone Oncology, Journal of Bone Oncology, 8, 18 - 22, Sep. 01 2017 , Refereed
    Summary:Background Bone represents one of the most common sites to which breast cancer cells metastasize. Patients experience skeletal related adverse events (pathological fractures, spinal cord compressions, and irradiation for deteriorated pain on bone) even during treatment with zoledronic acid (ZA). Therefore, we conducted a retrospective cohort study to investigate the predictive factors for symptomatic skeletal events (SSEs) in bone-metastasized breast cancer (b-MBC) patients. Methods We retrospectively collected data on b-MBC patients treated with ZA. Patient characteristics, including age, subtype, the presence of non-bone lesions, the presence of multiple bone metastases at the commencement of ZA therapy, duration of ZA therapy, the time interval between breast cancer diagnosis and the initiation of ZA therapy, and type of systemic therapy, presence of previous SSE were analyzed using multivariable logistic regression analysis. Results The medical records of 183 patients were reviewed and 176 eligible patients were analyzed. The median age was 59 (range, 30–87) years. Eighty-seven patients were aged ≥60 years and 89 patients were aged < 60 years. The proportions of patients with estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2-positive disease were 81.8%, 63.1%, and 17.6%, respectively. Fifty-three patients had bone-only MBC at the commencement of ZA therapy. SSEs were observed in 42 patients. In the multivariable logistic regression analysis, bone-only MBC but not a breast cancer subtype was an independent risk factor for an SSE during ZA therapy (odds ratio: 3.878, 95% confidence interval: 1.647–9.481 p = 0.002). Conclusions Bone-only MBC patients are more likely to experience an SSE even after treatment with ZA.
  • Increased risk of SSEs in bone-only metastatic breast cancer patients treated with zoledronic acid., Masashi Yanae, Shinichiro Fujimoto, Kaori Tane, Maki Tanioka, Kimiko Fujiwara, Masanobu Tsubaki, Yuzuru Yamazoe, Yoshiyuki Morishima, Yasutaka Chiba, Shintaro Takao, Yoshifumi Komoike, Junji Tsurutani, Kazuhiko Nakagawa, Shozo Nishida, Journal of bone oncology, Journal of bone oncology, 8, 18 - 22, Sep. 2017 , Refereed
    Summary:BACKGROUND: Bone represents one of the most common sites to which breast cancer cells metastasize. Patients experience skeletal related adverse events (pathological fractures, spinal cord compressions, and irradiation for deteriorated pain on bone) even during treatment with zoledronic acid (ZA). Therefore, we conducted a retrospective cohort study to investigate the predictive factors for symptomatic skeletal events (SSEs) in bone-metastasized breast cancer (b-MBC) patients. METHODS: We retrospectively collected data on b-MBC patients treated with ZA. Patient characteristics, including age, subtype, the presence of non-bone lesions, the presence of multiple bone metastases at the commencement of ZA therapy, duration of ZA therapy, the time interval between breast cancer diagnosis and the initiation of ZA therapy, and type of systemic therapy, presence of previous SSE were analyzed using multivariable logistic regression analysis. RESULTS: The medical records of 183 patients were reviewed and 176 eligible patients were analyzed. The median age was 59 (range, 30-87) years. Eighty-seven patients were aged ≥60 years and 89 patients were aged < 60 years. The proportions of patients with estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2-positive disease were 81.8%, 63.1%, and 17.6%, respectively. Fifty-three patients had bone-only MBC at the commencement of ZA therapy. SSEs were observed in 42 patients. In the multivariable logistic regression analysis, bone-only MBC but not a breast cancer subtype was an independent risk factor for an SSE during ZA therapy (odds ratio: 3.878, 95% confidence interval: 1.647-9.481; p = 0.002). CONCLUSIONS: Bone-only MBC patients are more likely to experience an SSE even after treatment with ZA.
  • Contributions of MET activation to BCR-ABL1 tyrosine kinase inhibitor resistance in chronic myeloid leukemia cells., Masanobu Tsubaki, Tomoya Takeda, Toshiki Kino, Kazuko Sakai, Tatsuki Itoh, Motohiro Imano, Takashi Nakayama, Kazuto Nishio, Takao Satou, Shozo Nishida, Oncotarget, Oncotarget, 8(24), 38717 - 38730, Jun. 13 2017 , Refereed
    Summary:Resistance to the breakpoint cluster region-abelson 1 (BCR-ABL1) tyrosine kinase inhibitor (TKI) imatinib poses a major problem when treating chronic myeloid leukemia (CML). Imatinib resistance often results from a secondary mutation in BCR-ABL1. However, in the absence of a mutation in BCR-ABL1, the basis of BCR-ABL1-independent resistance must be elucidated. To gain insight into the mechanisms of BCR-ABL1-independent imatinib resistance, we performed an array-based comparative genomic hybridization. We identified various resistance-related genes, and focused on MET. Treatment with a MET inhibitor resensitized K562/IR cells to BCR-ABL1 TKIs. Combined treatment of K562/IR cells with imatinib and a MET inhibitor suppressed extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK) activation, but did not affect AKT activation. Our findings implicate the MET/ERK and MET/JNK pathways in conferring resistance to imatinib, providing new insights into the mechanisms of BCR-ABL1 TKI resistance in CML.
  • The sensitivity of head and neck carcinoma cells to statins is related to the expression of their Ras expression status, and statin-induced apoptosis is mediated via suppression of the Ras/ERK and Ras/mTOR pathways, Masanobu Tsubaki, Daichiro Fujiwara, Tomoya Takeda, Toshiki Kino, Yoshika Tomonari, Tatsuki Itoh, Motohiro Imano, Takao Satou, Katsuhiko Sakaguchi, Shozo Nishida, CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, 44(2), 222 - 234, Feb. 2017 , Refereed
    Summary:Statins induce apoptosis of tumour cells by inhibiting the prenylation of small G-proteins. However, the details of the apoptosis-inducing mechanisms remain poorly understood. The present study showed that the induction of apoptosis by statins in four different human head and neck squamous cell carcinoma ( HNSCC) cell lines, HSC-3, HEp-2, Ca9-22, and SAS cells was mediated by increased caspase-3 activity. Statins induced apoptosis by the suppression of geranylgeranyl pyrophosphate biosynthesis. Furthermore, statins decreased the levels of phosphorylated ERK and mTOR by inhibiting the membrane localization of Ras and enhancing Bim expression in HSC-3 and HEp-2 cells. We also found that in all the cell types analyzed, the IC50 values for fluvastatin and simvastatin were highest in HEp-2 cells. In addition, HSC-3, Ca9-22, and SAS cells had higher Ras expression and membrane localization, higher activation of ERK1/2 and mTOR, and lower levels of Bim expression than HEp-2 cells. Our results indicate that statins induce apoptosis by increasing the activation of caspase-3 and by enhancing Bim expression through inhibition of the Ras/ERK and Ras/mTOR pathways. Furthermore, the sensitivity of HNSCC cells to statin treatment was closely related to Ras expression and prenylation levels, indicating that statins may act more effectively against tumours with high Ras expression and Ras-variability. Therefore, our findings support the use of statins as potential anticancer agents.
  • [Influence of Next-Day Administration of Pegfilgrastim after FEC100 Chemotherapy in Japanese with Breast Cancer on Neutrophil Count]., Daichiro Fujiwara, Keiji Mashimo, Kayo Kimura, Akihiro Noda, Kazuo Taki, Hiroshi Yoshibayashi, Tomoya Takeda, Masanobu Tsubaki, Shozo Nishida, Katsuhiko Sakaguchi, Gan to kagaku ryoho. Cancer & chemotherapy, Gan to kagaku ryoho. Cancer & chemotherapy, 44(2), 149 - 152, Feb. 2017 , Refereed
    Summary:Febrile neutropenia(FN)is one of the serious treatment-related toxicities after FEC100(5-fluorouracil 500mg/m2, epiru- bicin 100mg/m2, cyclophosphamide 500 mg/m2)chemotherapy for breast cancer. Granulocyte-colony stimulating factor(GCSF) is used as a support therapy for FN. Thus, we evaluated retrospectively the safety of administering pegfilgrastim the day after FEC100 chemotherapy in Japanese patients with breast cancer as compared with lenograstim. Grade 3 or 4 neutropenia was observed in 91.7% patients after pegfilgrastim administration and in 63.2% after lenograstim. The incidence rate of FN was 7.0%after pegfilgrastim administration and 9.7%after lenograstim, a difference that was not significantly different(p= 0.741). The mean relative dose intensity was good at 0.98 for pegfilgrastim and 0.97 for lenograstim. In conclusion, pegfilgrastim is not inferior to lenograstim in the incidence of FN. However, we do not recommend administering pegfilgrastim on the day after FEC100 therapy because it causes more severe neutropenia and has a high risk of FN. The timing of administration of pegfilgrastim in FEC100 therapy requires further study.
  • The sensitivity of head and neck carcinoma cells to statins is related to the expression of their Ras expression status, and statin-induced apoptosis is mediated via suppression of the Ras/ERK and Ras/mTOR pathways., Masanobu Tsubaki, Daichiro Fujiwara, Tomoya Takeda, Toshiki Kino, Yoshika Tomonari, Tatsuki Itoh, Motohiro Imano, Takao Satou, Katsuhiko Sakaguchi, Shozo Nishida, Clinical and experimental pharmacology & physiology, Clinical and experimental pharmacology & physiology, 44(2), 222 - 234, Feb. 2017 , Refereed
    Summary:Statins induce apoptosis of tumour cells by inhibiting the prenylation of small G-proteins. However, the details of the apoptosis-inducing mechanisms remain poorly understood. The present study showed that the induction of apoptosis by statins in four different human head and neck squamous cell carcinoma (HNSCC) cell lines, HSC-3, HEp-2, Ca9-22, and SAS cells was mediated by increased caspase-3 activity. Statins induced apoptosis by the suppression of geranylgeranyl pyrophosphate biosynthesis. Furthermore, statins decreased the levels of phosphorylated ERK and mTOR by inhibiting the membrane localization of Ras and enhancing Bim expression in HSC-3 and HEp-2 cells. We also found that in all the cell types analyzed, the IC50 values for fluvastatin and simvastatin were highest in HEp-2 cells. In addition, HSC-3, Ca9-22, and SAS cells had higher Ras expression and membrane localization, higher activation of ERK1/2 and mTOR, and lower levels of Bim expression than HEp-2 cells. Our results indicate that statins induce apoptosis by increasing the activation of caspase-3 and by enhancing Bim expression through inhibition of the Ras/ERK and Ras/mTOR pathways. Furthermore, the sensitivity of HNSCC cells to statin treatment was closely related to Ras expression and prenylation levels, indicating that statins may act more effectively against tumours with high Ras expression and Ras-variability. Therefore, our findings support the use of statins as potential anticancer agents.
  • Exploration of Molecular Targets in the Development of New Therapeutics Aimed at Overcoming Multidrug Resistance., Shozo Nishida, Masanobu Tsubaki, Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan, Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan, 137(2), 145 - 149, 2017 , Refereed
    Summary:Multidrug resistance (MDR) in cancer is a major problem in clinical settings: MDR correlates with a patient's poor prognosis and decreased quality of life. Recently, MDR was found to be involved in various signal pathways, so the inhibition of signal molecules by molecular targeting drugs may help overcome MDR. In addition, the acquisition of MDR is shown to be associated with the overexpression of drug efflux pumps such as P-glycoprotein (MDR1), which in turn affects the regulation of the expression of cell survival factors, B-cell leukemia protein 2 (Bcl-2) family proteins, etc. We analyzed the mechanisms of MDR in hematopoietic malignancies, and showed that the activation of signaling molecules regulated the expression of drug efflux pumps and cell survival factors, thus suggesting that molecular targeting drugs are potentially useful as anti-MDR agents. In this review, I focus on recent advancements in understanding the mechanisms of MDR with respect to hematopoietic malignancies: (1) exploration of molecular targets for overcoming MDR in anti-cancer drug-resistant cell lines, (2) the mechanism of drug resistance through the cytokine autocrine loop, and (3) cell-cell interaction with bone marrow stromal cells, along with the application of molecular targeting drugs for overcoming MDR.
  • Mangiferin, a novel nuclear factor kappa B-inducing kinase inhibitor, suppresses metastasis and tumor growth in a mouse metastatic melanoma model., Tomoya Takeda, Masanobu Tsubaki, Kotaro Sakamoto, Eri Ichimura, Aya Enomoto, Yuri Suzuki, Tatsuki Itoh, Motohiro Imano, Genzoh Tanabe, Osamu Muraoka, Hideaki Matsuda, Takao Satou, Shozo Nishida, Toxicology and applied pharmacology, Toxicology and applied pharmacology, 306, 105 - 12, Sep. 01 2016 , Refereed
    Summary:Advanced metastatic melanoma, one of the most aggressive malignancies, is currently without reliable therapy. Therefore, new therapies are urgently needed. Mangiferin is a naturally occurring glucosylxanthone and exerts many beneficial biological activities. However, the effect of mangiferin on metastasis and tumor growth of metastatic melanoma remains unclear. In this study, we evaluated the effect of mangiferin on metastasis and tumor growth in a mouse metastatic melanoma model. We found that mangiferin inhibited spontaneous metastasis and tumor growth. Furthermore, mangiferin suppressed the nuclear translocation of nuclear factor kappa B (NF-κB) and expression of phosphorylated NF-κB-inducing kinase (NIK), inhibitor of kappa B kinase (IKK), and inhibitor of kappa B (IκB) and increases the expression of IκB protein in vivo. In addition, we found that mangiferin inhibited the expression of matrix metalloproteinases (MMPs) and very late antigens (VLAs) in vivo. Mangiferin treatment also increased the expression of cleaved caspase-3, cleaved Poly ADP ribose polymerase-1 (PARP-1), p53 upregulated modulator of apoptosis (PUMA), p53, and phosphorylated p53 proteins, and decreased the expression of Survivin and Bcl-associated X (Bcl-xL) proteins in vivo. These results indicate that mangiferin selectivity suppresses the NF-κB pathway via inhibition of NIK activation, thereby inhibiting metastasis and tumor growth. Importantly, the number of reported NIK selective inhibitors is limited. Taken together, our data suggest that mangiferin may be a potential therapeutic agent with a new mechanism of targeting NIK for the treatment of metastatic melanoma.
  • RANK-RANKL interactions are involved in cell adhesion-mediated drug resistance in multiple myeloma cell lines., Masanobu Tsubaki, Tomoya Takeda, Misako Yoshizumi, Emi Ueda, Tatsuki Itoh, Motohiro Imano, Takao Satou, Shozo Nishida, Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, 37(7), 9099 - 110, Jul. 2016 , Refereed
    Summary:Interaction between multiple myeloma (MM) cells and the bone marrow microenvironment plays a critical role in MM pathogenesis and the development of drug resistance. Recently, it has been reported that MM cells express the receptor activator of nuclear factor-κB (NF-κB) (RANK). However, the role of the RANK/RANK ligand (RANKL) system in drug resistance remains unclear. In this study, we demonstrated a novel function of the RANK/RANKL system in promoting drug resistance in MM. We found that RANKL treatment induced drug resistance in RANK-expressing but not RANK-negative cell lines. RANKL stimulation of RANK-expressing cells increased multidrug resistance protein 1 (MDR1), breast cancer resistance protein (BCRP), and lung resistance protein 1 (LRP1) expression and decreased Bim expression through various signaling molecules. RNA silencing of Bim expression induced drug resistance, but the RANKL-mediated drug resistance could not be overcome through the RNA silencing of MDR1, BCRP, and LRP1 expression. These results indicate that the RANK/RANKL system induces chemoresistance through the activation of multiple signal transduction pathways and by decreasing Bim expression in RANK-positive MM cells. These findings may prove to be useful in the development of cell adhesion-mediated drug resistance inhibitors in RANK-positive MM cells.
  • Mangiferin induces apoptosis in multiple myeloma cell lines by suppressing the activation of nuclear factor kappa B-inducing kinase., Tomoya Takeda, Masanobu Tsubaki, Toshiki Kino, Misa Yamagishi, Megumi Iida, Tatsuki Itoh, Motohiro Imano, Genzoh Tanabe, Osamu Muraoka, Takao Satou, Shozo Nishida, Chemico-biological interactions, Chemico-biological interactions, 251, 26 - 33, May 05 2016 , Refereed
    Summary:Mangiferin is a naturally occurring glucosyl xanthone, which induces apoptosis in various cancer cells. However, the molecular mechanism underlying mangiferin-induced apoptosis has not been clarified thus far. Therefore, we examined the molecular mechanism underlying mangiferin-induced apoptosis in multiple myeloma (MM) cell lines. We found that mangiferin decreased the viability of MM cell lines in a concentration-dependent manner. We also observed an increased number of apoptotic cells, caspase-3 activation, and a decrease in the mitochondrial membrane potential. In addition, mangiferin inhibited the nuclear translocation of nuclear factor kappa B (NF-κB) and expression of phosphorylated inhibitor kappa B (IκB) and increased the expression of IκB protein, whereas no changes were observed in the phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal protein kinase 1/2 (JNK1/2), and mammalian target of rapamycin (mTOR). The molecular mechanism responsible for mangiferin-induced inhibition of nuclear translocation of NF-κB was a decrease in the expression of phosphorylated NF-κB-inducing kinase (NIK). Moreover, mangiferin decreased the expression of X-linked inhibitor of apoptosis protein (XIAP), survivin, and Bcl-xL proteins. Knockdown of NIK expression showed results similar to those observed with mangiferin treatment. Our results suggest that mangiferin induces apoptosis through the inhibition of nuclear translocation of NF-κB by suppressing NIK activation in MM cell lines. Our results provide a new insight into the molecular mechanism of mangiferin-induced apoptosis. Importantly, since the number of reported NIK inhibitors is limited, mangiferin, which targets NIK, may be a potential anticancer agent for the treatment of MM.
  • PKN3 is the major regulator of angiogenesis and tumor metastasis in mice., Hideyuki Mukai, Aiko Muramatsu, Rana Mashud, Koji Kubouchi, Sho Tsujimoto, Tsunaki Hongu, Yasunori Kanaho, Masanobu Tsubaki, Shozo Nishida, Go Shioi, Sally Danno, Mona Mehruba, Ryosuke Satoh, Reiko Sugiura, Scientific reports, Scientific reports, 6, 18979 - 18979, Jan. 08 2016 , Refereed
    Summary:PKN, a conserved family member related to PKC, was the first protein kinase identified as a target of the small GTPase Rho. PKN is involved in various functions including cytoskeletal arrangement and cell adhesion. Furthermore, the enrichment of PKN3 mRNA in some cancer cell lines as well as its requirement in malignant prostate cell growth suggested its involvement in oncogenesis. Despite intensive research efforts, physiological as well as pathological roles of PKN3 in vivo remain elusive. Here, we generated mice with a targeted deletion of PKN3. The PKN3 knockout (KO) mice are viable and develop normally. However, the absence of PKN3 had an impact on angiogenesis as evidenced by marked suppressions of micro-vessel sprouting in ex vivo aortic ring assay and in vivo corneal pocket assay. Furthermore, the PKN3 KO mice exhibited an impaired lung metastasis of melanoma cells when administered from the tail vein. Importantly, PKN3 knock-down by small interfering RNA (siRNA) induced a glycosylation defect of cell-surface glycoproteins, including ICAM-1, integrin β1 and integrin α5 in HUVECs. Our data provide the first in vivo genetic demonstration that PKN3 plays critical roles in angiogenesis and tumor metastasis, and that defective maturation of cell surface glycoproteins might underlie these phenotypes.
  • PKC/MEK inhibitors suppress oxaliplatin-induced neuropathy and potentiate the antitumor effects., Masanobu Tsubaki, Tomoya Takeda, Tadahumi Tani, Hirotaka Shimaoka, Naohiro Suzuyama, Kotaro Sakamoto, Arisa Fujita, Naoki Ogawa, Tatsuki Itoh, Motohiro Imano, Yoshinori Funakami, Seiji Ichida, Takao Satou, Shozo Nishida, International journal of cancer, International journal of cancer, 137(1), 243 - 50, Jul. 01 2015 , Refereed
    Summary:Oxaliplatin is a key drug commonly used in colorectal cancer treatment. Despite high clinical efficacy, its therapeutic application is limited by common, dose-limiting occurrence of neuropathy. As usual symptomatic neuropathy treatments fail to improve the patients' condition, there is an urgent need to advance our understanding of the pathogenesis of neuropathy to propose effective therapy and ensure adequate pain management. Oxaliplatin-induced neuropathy was recently reported to be associated with protein kinase C (PKC) activation. It is unclear, however, whether PKC inhibition can prevent neuropathy. In our current studies, we found that a PKC inhibitor, tamoxifen, inhibited oxaliplatin-induced neuropathy via the PKC/extracellular signal-regulated kinase (ERK)/c-Fos pathway in lumbar spinal cords (lumbar segments 4-6). Additionally, tamoxifen was shown to act in synergy with oxaliplatin to inhibit growth in tumor cells-implanted mice. Moreover, mitogen-activated protein kinase kinase (MEK) 1/2 inhibitor, PD0325901, suppressed oxaliplatin-induced neuropathy and enhanced oxaliplatin efficacy. Our results indicate that oxaliplatin-induced neuropathy is associated with PKC/ERK/c-Fos pathway in lumbar spinal cord. Additionally, we demonstrate that disruption of this pathway by PKC and MEK inhibitors suppresses oxaliplatin-induced neuropathy, thereby suggesting that PKC and MEK inhibitors may be therapeutically useful in preventing oxaliplatin-induced neuropathy and could aid in combination antitumor pharmacotherapy.
  • A phase II trial of perioperative chemotherapy involving a single intraperitoneal administration of paclitaxel followed by sequential S-1 plus intravenous paclitaxel for serosa-positive gastric cancer., Ying-Feng Peng, Motohiro Imano, Tatsuki Itoh, Takao Satoh, Yasutaka Chiba, Haruhiko Imamoto, Masahiro Tsubaki, Shozo Nishida, Takushi Yasuda, Hiroshi Furukawa, Journal of surgical oncology, Journal of surgical oncology, 111(8), 1041 - 6, Jun. 2015 , Refereed
    Summary:BACKGROUND AND OBJECTIVES: We carried out a phase II trial to evaluate the feasibility, efficacy, and tolerability of perioperative chemotherapy including single intraperitoneal(IP) administration of paclitaxel(PTX) followed by intravenous(IV) administrations of PTX with S-1 in a neoadjuvant setting for serosa-positive gastric cancer. METHODS: Patients with cT4a gastric cancer were enrolled. A laparoscopic survey was performed before study inclusion for the confirmation of serosal invasion, negative lavage cytology, and negative peritoneal metastasis. IP PTX (80 mg/m(2)) was administered, followed by systemic chemotherapy. Surgery was performed after the completion of chemotherapy. The primary endpoint was the treatment completion rate. RESULTS: 37 patients were recruited. The treatment completion rate was 67.6% (25/37; 90% CI, 52.8-80.1%), which was significantly higher than 50%; we set this as a threshold value (P = 2.4% [one-sided]). 14 patients had target lesions; of these, 10 showed a partial response (71.4%), three had stable disease (21.4%), and one had progressive disease(7.2%). The response rate was 71.4% (10/14). All patients underwent gastrectomy with D2 lymph node dissection. The 3- and 5-year OS rates were 78.0 and 74.9%, respectively. CONCLUSIONS: Perioperative chemotherapy including neoadjuvant IP PTX followed by sequential IV PTX with S-1 for serosa-positive gastric cancer is feasible, safe, and efficient.
  • Statins improve survival by inhibiting spontaneous metastasis and tumor growth in a mouse melanoma model., Masanobu Tsubaki, Tomoya Takeda, Toshiki Kino, Naoya Obata, Tatsuki Itoh, Motohiro Imano, Kenji Mashimo, Daichiro Fujiwara, Katsuhiko Sakaguchi, Takao Satou, Shozo Nishida, American journal of cancer research, American journal of cancer research, 5(10), 3186 - 97, 2015 , Refereed
    Summary:Metastatic melanoma is a life-threatening disease for which no effective treatment is currently available. In melanoma cells, Rho overexpression promotes invasion and metastasis. However, the effect of statins on spontaneous metastasis and tumor growth remains unclear. In the present study, we investigated the mechanism of statin-mediated tumor growth and metastasis inhibition in an in vivo model. We found that statins significantly inhibited spontaneous metastasis and tumor growth. Statins inhibited the mRNA expression and enzymatic activities of matrix metalloproteinases (MMPs) in vivo and also suppressed the mRNA and protein expression of very late antigens (VLAs). Moreover, statins inhibited the prenylation of Rho as well as the phosphorylation of LIM kinase, serum response factor (SRF), and c-Fos downstream of the Rho signaling pathway. In addition, statins enhanced p53, p21, and p27 expression and reduced phosphorylation of cyclin-dependent kinase and expression of cyclin D1 and E2. These results indicate that statins suppress Rho signaling pathways, thereby inhibiting tumor metastasis and growth. Furthermore, statins markedly improved the survival rate in a metastasis model, suggesting that statins have potential clinical applications for the treatment of metastatic cancers.
  • Nitrogen-containing bisphosphonates inhibit RANKL- and M-CSF-induced osteoclast formation through the inhibition of ERK1/2 and Akt activation., Masanobu Tsubaki, Makiko Komai, Tatsuki Itoh, Motohiro Imano, Kotaro Sakamoto, Hirotaka Shimaoka, Tomoya Takeda, Naoki Ogawa, Kenji Mashimo, Daiichiro Fujiwara, Junji Mukai, Katsuhiko Sakaguchi, Takao Satou, Shozo Nishida, Journal of biomedical science, Journal of biomedical science, 21(1), 10 - 10, Feb. 03 2014 , Refereed
    Summary:BACKGROUND: Bisphosphonates are an important class of antiresorptive drugs used in the treatment of metabolic bone diseases. Recent studies have shown that nitrogen-containing bisphosphonates induced apoptosis in rabbit osteoclasts and prevented prenylated small GTPase. However, whether bisphosphonates inhibit osteoclast formation has not been determined. In the present study, we investigated the inhibitory effect of minodronate and alendronate on the osteoclast formation and clarified the mechanism involved in a mouse macrophage-like cell lines C7 and RAW264.7. RESULTS: It was found that minodronate and alendronate inhibited the osteoclast formation of C7 cells induced by receptor activator of NF-κB ligand and macrophage colony stimulating factor, which are inhibited by the suppression of geranylgeranyl pyrophosphate (GGPP) biosynthesis. It was also found that minodronate and alendronate inhibited the osteoclast formation of RAW264.7 cells induced by receptor activator of NF-κB ligand. Furthermore, minodronate and alendornate decreased phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt; similarly, U0126, a mitogen protein kinase kinase 1/2 (MEK1/2) inhibitor, and LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, inhibited osteoclast formation. CONCLUSIONS: This indicates that minodronate and alendronate inhibit GGPP biosynthesis in the mevalonate pathway and then signal transduction in the MEK/ERK and PI3K/Akt pathways, thereby inhibiting osteoclast formation. These results suggest a novel effect of bisphosphonates that could be effective in the treatment of bone metabolic diseases, such as osteoporosis.
  • Risk factors for cisplatin-induced nephrotoxicity and potential of magnesium supplementation for renal protection., Yasuhiro Kidera, Hisato Kawakami, Tsutomu Sakiyama, Kunio Okamoto, Kaoru Tanaka, Masayuki Takeda, Hiroyasu Kaneda, Shin-ichi Nishina, Junji Tsurutani, Kimiko Fujiwara, Morihiro Nomura, Yuzuru Yamazoe, Yasutaka Chiba, Shozo Nishida, Takao Tamura, Kazuhiko Nakagawa, PloS one, PloS one, 9(7), e101902, 2014 , Refereed
    Summary:BACKGROUND: Nephrotoxicity remains a problem for patients who receive cisplatin chemotherapy. We retrospectively evaluated potential risk factors for cisplatin-induced nephrotoxicity as well as the potential impact of intravenous magnesium supplementation on such toxicity. PATIENTS AND METHODS: We reviewed clinical data for 401 patients who underwent chemotherapy including a high dose (≥60 mg/m2) of cisplatin in the first-line setting. Nephrotoxicity was defined as an increase in the serum creatinine concentration of at least grade 2 during the first course of cisplatin chemotherapy, as assessed on the basis of National Cancer Institute Common Terminology Criteria for Adverse Events version 4.0. The severity of nephrotoxicity was evaluated on the basis of the mean change in the serum creatinine level. Magnesium was administered intravenously to 67 patients (17%). RESULTS: Cisplatin-induced nephrotoxicity was observed in 127 patients (32%). Multivariable analysis revealed that an Eastern Cooperative Oncology Group performance status of 2 (risk ratio, 1.876; P = 0.004) and the regular use of nonsteroidal anti-inflammatory drugs (NSAIDs) (risk ratio, 1.357; P = 0.047) were significantly associated with an increased risk for cisplatin nephrotoxicity, whereas intravenous magnesium supplementation was associated with a significantly reduced risk for such toxicity (risk ratio, 0.175; P = 0.0004). The development of hypomagnesemia during cisplatin treatment was significantly associated with a greater increase in serum creatinine level (P = 0.0025). Magnesium supplementation therapy was also associated with a significantly reduced severity of renal toxicity (P = 0.012). CONCLUSIONS: A relatively poor performance status and the regular use of NSAIDs were significantly associated with cisplatin-induced nephrotoxicity, although the latter association was marginal. Our findings also suggest that the ability of magnesium supplementation to protect against the renal toxicity of cisplatin warrants further investigation in a prospective trial.
  • By inhibiting Src, verapamil and dasatinib overcome multidrug resistance via increased expression of Bim and decreased expressions of MDR1 and survivin in human multidrug-resistant myeloma cells., Masanobu Tsubaki, Makiko Komai, Tatsuki Itoh, Motohiro Imano, Kotaro Sakamoto, Hirotaka Shimaoka, Tomoya Takeda, Naoki Ogawa, Kenji Mashimo, Daiichiro Fujiwara, Junji Mukai, Katsuhiko Sakaguchi, Takao Satou, Shozo Nishida, Leukemia research, Leukemia research, 38(1), 121 - 30, Jan. 2014 , Refereed
    Summary:The calcium channel blocker verapamil inhibits the transport function of multidrug resistance protein 1 (MDR1). Although verapamil acts to reverse MDR in cancer cells, the underlying mechanism remains unclear. In the present study, we investigated the mechanism of reversing MDR by verapamil in anti-cancer drug-resistant multiple myeloma (MM) cell lines. We found that verapamil suppresses MDR1 and survivin expressions and increases Bim expression via suppression of Src activation. Furthermore, dasatinib reversed the drug-resistance of the drug-resistant cell lines. These findings suggest that Src inhibitors are potentially useful as an anti-MDR agent for the treatment of malignant tumor cells.
  • High expression of epithelial cellular adhesion molecule in peritoneal metastasis of gastric cancer., Motohiro Imano, Tatsuki Itoh, Takao Satou, Atsushi Yasuda, Kohei Nishiki, Hiroaki Kato, Osamu Shiraishi, Ying-Feng Peng, Masayuki Shinkai, Masahiro Tsubaki, Takushi Yasuda, Haruhiko Imamoto, Shozo Nishida, Yoshifumi Takeyama, Hiroshi Furkawa, Kiyokata Okuno, Hitoshi Shiozaki, Targeted oncology, Targeted oncology, 8(4), 231 - 5, Dec. 2013 , Refereed
    Summary:Intraperitoneally administrated epithelial cellular adhesion molecule (EpCAM) monoclonal antibody is a therapeutic agent in patients with malignant effusion in several types of carcinoma. However, the role of EpCAM in peritoneal metastasis (PM) lesions and primary lesions of gastric cancer (GC) is still unclear. Therefore, in this study, we investigated EpCAM expression in GC patients with PM. We investigated the expression of EpCAM in 35PM lesions and 104 biopsy samples as primary lesions. Immunohistochemical staining was performed using the Ventana Benchmark XT (Roche Diagnostics) system. EpCAM expression was evaluated by calculating the total immunostaining score, which is the product of the proportion score and the intensity score. Overexpression was defined as a total score greater than 4. All PM specimens showed overexpression of EpCAM, and GC cells in both the surface layer and the deep layer of the PM showed a high expression of EpCAM. Meanwhile, in the biopsy sample, the expression of EpCAM ranged from none to strong. The EpCAM score results for PM specimens and biopsy samples were 11.0 ± 2.0 and 6.9 ± 3.9, respectively. The difference between the scores was statistically significant (P < 0.05). The intraperitoneally administrated EpCAM antibody might have a anti-cancer effect in PM lesions of GC. Additionally, it can be assumed that only GC cells which express a high level of EpCAM might metastasize to the peritoneum.
  • Inhibition of the tumour necrosis factor-alpha autocrine loop enhances the sensitivity of multiple myeloma cells to anticancer drugs., Masanobu Tsubaki, Makiko Komai, Tatsuki Itoh, Motohiro Imano, Kotaro Sakamoto, Hirotaka Shimaoka, Naoki Ogawa, Kenji Mashimo, Daichiro Fujiwara, Tomoya Takeda, Junji Mukai, Katsuhiko Sakaguchi, Takao Satou, Shozo Nishida, European journal of cancer (Oxford, England : 1990), European journal of cancer (Oxford, England : 1990), 49(17), 3708 - 17, Nov. 2013 , Refereed
    Summary:Several autocrine soluble factors, including macrophage inflammatory protein-1α and tumour necrosis factor-alpha (TNF-α), promote the survival and growth of multiple myeloma (MM) cells. We hypothesised that inhibition of the TNF-α autocrine loop may enhance the cytotoxic effect of anticancer drugs in MM cell lines. In the present study, a TNF-α-neutralizing antibody suppressed cell proliferation and enhanced the cytotoxic effect of anticancer drugs on MM cells. In addition, combination treatment with the TNF-α-neutralizing antibody and the chemotherapy agent melphalan inhibited nuclear factor κB (NF-κB) p65 nuclear translocation and mammalian target of rapamycin (mTOR) activation and upregulated the expression of Bax and Bim. Treatment of ARH-77 cells with the NF-κB inhibitor dimethyl fumarate or the mTOR inhibitor rapamycin suppressed NF-κB p65 nuclear translocation and enhanced the cytotoxic effect of melphalan. Furthermore, infliximab, a monoclonal antibody against TNF-α, also enhanced the cytotoxic effect of anticancer drugs in ARH-77 cells. These results indicated that TNF-α-neutralizing antibodies or infliximab enhanced the cytotoxic effect of anticancer drugs by suppressing the TNF receptor/mTOR/NF-κB pathways. The inhibition of TNF-α may thus provide a new therapeutic approach to control tumour progression and bone destruction in MM patients.
  • [A search for the risk factors for hiccups and evaluation of antiemetic therapy in CDDP-based chemotherapy, using cluster analysis]., Hajime Asano, Mizuki Watanabe, Akinori Kawaguchi, Masashi Yanae, Yoshinori Funakami, Tetsuyuki Wada, Sumio Matzno, Yuzuru Yamazoe, Shozo Nishida, Seiji Ichida, Gan to kagaku ryoho. Cancer & chemotherapy, Gan to kagaku ryoho. Cancer & chemotherapy, 40(8), 1031 - 6, Aug. 2013 , Refereed
    Summary:Hiccups are often observed in patients treated with cisplatin(CDDP)-based chemotherapy. It has been reported that gender and specific dosages of CDDP and antiemetic drugs(e.g., dexamethasone and 5-HT3 receptor antagonist)using standard therapy are major risk factors in the onset of hiccups. Recently, aprepitant has been added to the antiemetic therapy in CDDP-based chemotherapy. However, it is not known how the onset of hiccups takes place in antiemetic therapy including aprepitant according to the guideline. In this study, we used cluster analysis to classify 229 patients treated with CDDP-based chemotherapy, to investigate the effect of antiemetic therapy on the onset of hiccups and chemotherapy-induced nausea and vomiting(CINV). Our analysis indicated that aprepitant was not a major risk factor for the onset of hiccups in the high CDDP dose group(≥70 mg/m(2)). However, an effect of antiemesis was confirmed in the standard therapy with aprepitant. In conclusion, we suggest that aprepitant is effective for CINV, without causing the onset of hiccups in patients treated with high-dose CDDP-based chemotherapy.
  • Neuroprotective effect of (-)-epigallocatechin-3-gallate in rats when administered pre- or post-traumatic brain injury., Tatsuki Itoh, Masaki Tabuchi, Nobuyuki Mizuguchi, Motohiro Imano, Masahiro Tsubaki, Shozo Nishida, Shigeo Hashimoto, Kazuhiko Matsuo, Takashi Nakayama, Akihiko Ito, Hiroshi Munakata, Takao Satou, Journal of neural transmission (Vienna, Austria : 1996), Journal of neural transmission (Vienna, Austria : 1996), 120(5), 767 - 83, May 2013 , Refereed
    Summary:Our previous study indicated that consuming (-)-epigallocatechin gallate (EGCG) before or after traumatic brain injury (TBI) eliminated free radical generation in rats, resulting in inhibition of neuronal degeneration and apoptotic death, and improvement of cognitive impairment. Here we investigated the effects of administering EGCG at various times pre- and post-TBI on cerebral function and morphology. Wistar rats were divided into five groups and were allowed access to (1) normal drinking water, (2) EGCG pre-TBI, (3) EGCG pre- and post-TBI, (4) EGCG post-TBI, and (5) sham-operated group with access to normal drinking water. TBI was induced with a pneumatic controlled injury device at 10 weeks of age. Immunohistochemistry and lipid peroxidation studies revealed that at 1, 3, and 7 days post-TBI, the number of 8-Hydroxy-2'-deoxyguanosine-, 4-Hydroxy-2-nonenal- and single-stranded DNA (ssDNA)-positive cells, and levels of malondialdehyde around the damaged area were significantly decreased in all EGCG treatment groups compared with the water group (P < 0.05). Although there was a significant increase in the number of surviving neurons after TBI in each EGCG treatment group compared with the water group (P < 0.05), significant improvement of cognitive impairment after TBI was only observed in the groups with continuous and post-TBI access to EGCG (P < 0.05). These results indicate that EGCG inhibits free radical-induced neuronal degeneration and apoptotic death around the area damaged by TBI. Importantly, continuous and post-TBI access to EGCG improved cerebral function following TBI. In summary, consumption of green tea may be an effective therapy for TBI patients.
  • Establishment of a novel model of peritoneal carcinomatosis of the peritoneal extension type., Motohiro Imano, Tatsuki Itoh, Takao Satou, Akira Kido, Masahiro Tsubaki, Atsushi Yasuda, Hiroaki Kato, Haruhiko Imamoto, Shozo Nishida, Hiroshi Furukawa, Yoshifumi Takeyama, Kiyokata Okuno, Hitoshi Shiozaki, Anticancer research, Anticancer research, 33(4), 1439 - 46, Apr. 2013 , Refereed
    Summary:AIM: Patients with scirrhous carcinoma of the gastrointestinal tract frequently develop peritoneal carcinomatosis-particularly of the peritoneal extension type (PET), which has a bad prognosis. We developed a novel animal model, suitable for testing treatments for PET. MATERIAL AND METHODS: In order to develop the model, we scraped the entire peritoneum of Fischer 344 rats with sterile cotton swabs and injected 1 × 10(6) cells of the RCN-9 cell type into the peritoneal cavity. RESULTS: In the novel experimental model, RCN-9 cells adhered only to the exposed basement membrane. The submesothelial layer and fibroblasts in the submesothelial layer grew and increased to a maximum at day 7, then decreased during late-phase peritoneal carcinomatosis. At day 14, RCN-9 cells coated the peritoneum in a manner similar to PET. CONCLUSION: We successfully established a novel animal model of peritoneal carcinomatosis that mimics clinicopathological features of PET. Fibroblasts in the submesothelial layer potentially play an important role in peritoneal carcinomatosis.
  • Association between occupational exposure and control measures for antineoplastic drugs in a pharmacy of a hospital., Jin Yoshida, Shigeki Koda, Shozo Nishida, Hiroyuki Nakano, Genshin Tei, Shinji Kumagai, The Annals of occupational hygiene, The Annals of occupational hygiene, 57(2), 251 - 60, Mar. 2013 , Refereed
    Summary:OBJECTIVES: To investigate the association between occupational contamination and exposure levels to antineoplastic drugs and the application of control measures in a hospital work environment. METHODS: Wipe samples of equipments were collected at a hospital in Osaka Prefecture, Japan, from 2007 to 2011. These samples were subjected to measurements of cyclophosphamide (CP), gemcitabine (GEM), platinum-containing drugs (Pt), and fluorouracil (5FU). Additionally, 24-h urine samples were collected from pharmacists who handled antineoplastic drugs, which were analyzed for CP and alpha-fluoro-beta-alanine (AFBA). The application of control measures was scored according to a checklist, which consisted of the following five items: safety equipment and maintenance, training and documentation, devices for safe handling, personal protective equipment, and emergency care. The aim was to obtain a score of 80%. RESULTS: The median CP, GEM, and 5FU concentrations of all wipe samples were significantly lower during the period when the mean score was >80% (attainment period) versus when the mean score was ≤80% (nonattainment period; all P < 0.001, Mann-Whitney's U-test). Additionally, the median urinary CP and AFBA concentrations of pharmacists during the attainment period tended to be lower than that of those during the nonattainment period (P = 0.061 and 0.061, respectively, using Mann-Whitney's U-test). CONCLUSIONS: Contamination and levels of exposure to antineoplastic drugs decreased with a score higher than 80%. The scores of the items on the checklist appeared to adequately reflect the condition of the control measures, as increases in all five items were associated with reductions in the contamination by and levels of exposure to all drugs.
  • Appearance of neural stem cells around the damaged area following traumatic brain injury in aged rats., Tatsuki Itoh, Motohiro Imano, Shozo Nishida, Masahiro Tsubaki, Takashi Nakayama, Nobuyuki Mizuguchi, Shigeaki Yamanaka, Masaki Tabuchi, Hiroshi Munakata, Shigeo Hashimoto, Akihiko Ito, Takao Satou, Journal of neural transmission (Vienna, Austria : 1996), Journal of neural transmission (Vienna, Austria : 1996), 120(3), 361 - 74, Mar. 2013 , Refereed
    Summary:We have previously reported free radical production after traumatic brain injury (TBI), which induces neural stem cell (NSC) degeneration and death. However, the effects of aging on NSC proliferation around the damaged area following TBI have not been investigated. Therefore, in this study, we used 10-week (young group) and 24-month-old (aged group) rat TBI models to investigate the effects of aging on NSC proliferation around damaged tissue using immunohistochemical and ex vivo techniques. Young and aged rats received TBI. At 1, 3 and 7 days after TBI, immunohistochemical and lipid peroxidation studies were performed. Immunohistochemistry revealed that the number of nestin-positive cells around the damaged area after TBI in the aged group decreased significantly when compared with those in the young group (P < 0.01). However, the number of 8-hydroxy-2'-deoxyguanosine-, 4-hydroxy-2-nonenal- and single-stranded DNA (ssDNA)-positive cells and the level of peroxidation around the damaged area after TBI significantly increased in the aged group, compared with those in the young group (P < 0.01). Furthermore, almost all ssDNA-positive cells in young and aged groups co-localized with NeuN and nestin staining. Ex vivo studies revealed that neurospheres, which differentiated into neurons and glia in culture, could only be isolated from injured brain tissue in young and aged groups at 3 days after TBI. These results indicate that, although there were fewer NSCs that have the potential to differentiate into neurons and glia, these NSCs escaped free radical-induced degeneration around the damaged area after TBI in the aged rat brain.
  • Nitrogen-containing bisphosphonates induce apoptosis of hematopoietic tumor cells via inhibition of Ras signaling pathways and Bim-mediated activation of the intrinsic apoptotic pathway., Masanobu Tsubaki, Tatsuki Itoh, Takao Satou, Motohiro Imano, Makiko Komai, Naoki Ogawa, Junji Mukai, Shozo Nishida, Biochemical pharmacology, Biochemical pharmacology, 85(2), 163 - 72, Jan. 15 2013 , Refereed
    Summary:Nitrogen-containing bisphosphonates (N-BPs) induce apoptosis in tumor cells by inhibiting the prenylation of small G-proteins. However, the details of the apoptosis-inducing mechanism remain obscure. The present study showed that the induction of apoptosis by N-BPs in hematopoietic tumor cells is mediated by mitochondrial apoptotic signaling pathways, which are activated by the suppression of geranylgeranyl pyrophosphate (GGPP) biosynthesis. Furthermore, N-BPs decreased the levels of phosphorylated extracellular signal-regulated kinase (ERK) and mTOR via suppression of Ras prenylation and enhanced Bim expression. The present results indicated that N-BPs induce apoptosis by decreasing the mitochondrial transmembrane potential, increasing the activation of caspase-9 and caspase-3, and enhancing Bim expression through inhibition of the Ras/MEK/ERK and Ras/mTOR pathways. The accumulation of N-BPs in bones suggests that they may act more effectively on tumors that have spread to bones or on Ras-variable tumors. This is the first study to show that the specific molecular pathways of N-BP-induced apoptosis.
  • Increased apoptotic neuronal cell death and cognitive impairment at early phase after traumatic brain injury in aged rats., Tatsuki Itoh, Motohiro Imano, Shozo Nishida, Masahiro Tsubaki, Nobuyuki Mizuguchi, Shigeo Hashimoto, Akihiko Ito, Takao Satou, Brain structure & function, Brain structure & function, 218(1), 209 - 20, Jan. 2013 , Refereed
    Summary:Progressive age-associated increases in cerebral dysfunction have been shown to occur following traumatic brain injury (TBI). Moreover, levels of neuronal mitochondrial antioxidant enzymes in the aged brain are reduced, resulting in free radical-induced cell death. It was hypothesized that cognitive impairment after TBI in the aged progresses to a greater degree than in younger individuals, and that damage involves neuronal degeneration and death by free radicals. In this study, we investigated the effects of free radicals on neuronal degeneration, cell death, and cognitive impairment in 10-week-old (young group) and 24-month-old rats (aged group) subjected to TBI. Young and aged rats received TBI with a pneumatic controlled injury device. At 1, 3 and 7 days after TBI, immunohistochemistry, lipid peroxidation and behavioral studies were performed. At 1, 3 and 7 days post-TBI, the number of 8-hydroxy-2'-deoxyguanosine-, 4-hydroxy-2-nonenal- and single-stranded DNA (ssDNA)-positive cells, and the levels of malondialdehyde around the damaged area after TBI significantly increased in the aged group when compared with the young group (P < 0.05). In addition, the majority of ssDNA-positive cells in both groups co-localized with neuronal cells around the damaged area. There was a significant decrease in the number of surviving neurons and an increase in cognitive impairment after TBI in the aged group when compared with the young group (P < 0.05). These results indicate that following TBI, high levels of free radicals are produced in the aged rat brain, which induces neuronal degeneration and apoptotic cell death around the damaged area, resulting in cognitive impairment.
  • Phase II study of single intraperitoneal chemotherapy followed by systemic chemotherapy for gastric cancer with peritoneal metastasis., Motohiro Imano, Atsushi Yasuda, Tatsuki Itoh, Takao Satou, Ying-Feng Peng, Hiroaki Kato, Masayuki Shinkai, Masahiro Tsubaki, Yasutaka Chiba, Takushi Yasuda, Haruhiko Imamoto, Shozo Nishida, Yoshifumi Takeyama, Kiyokata Okuno, Hiroshi Furukawa, Hitoshi Shiozaki, Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract, Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract, 16(12), 2190 - 6, Dec. 2012 , Refereed
    Summary:BACKGROUND: We conducted a phase II study involving a single administration of intraperitoneal chemotherapy with paclitaxel followed by sequential systemic chemotherapy with S-1+ paclitaxel for advanced gastric cancer patients with peritoneal metastasis. METHODS: Gastric cancer patients with peritoneal metastasis were enrolled. Paclitaxel (80 mg/m(2)) was administered intraperitoneally at staging laparoscopy. Within 7 days, patients received systemic chemotherapy with S-1 (80 mg/m(2)/day on days 1-14) plus paclitaxel (50 mg/m(2) on days 1 and 8), followed by 7-days rest. The responders to this chemotherapy underwent second-look laparoscopy, and gastrectomy with D2 lymph node dissection was performed in patients when the disappearance of peritoneal metastasis had been confirmed. The primary endpoint of the study was overall survival rate. RESULTS: Thirty-five patients were enrolled. All patients were confirmed as having localized peritoneal metastasis by staging laparoscopy. Eventually, gastrectomy was performed in 22 patients. The median survival time of the total patient population and those patients in which gastrectomy was performed was 21.3 and 29.8 months, respectively. The overall response rate was 65.7 % for all patients. The frequent grade 3/4 toxic effects included neutropenia and leukopenia. CONCLUSIONS: Sequential intraperitoneal and intravenous paclitaxel plus S-1 was well tolerated in gastric cancer patients with peritoneal metastasis.
  • Peritoneal metastatic lesions of gastric cancer exhibit low expression of human epidermal growth factor receptor 2., Motohiro Imano, Takao Satou, Tatsuki Itoh, Atsushi Yasuda, Hiroaki Kato, Masayuki Shinkai, Ying-Feng Peng, Masahiro Tsubaki, Takushi Yasuda, Haruhiko Imamoto, Shozo Nishida, Yoshifumi Takeyama, Kiyokata Okuno, Hitoshi Shiozaki, Targeted oncology, Targeted oncology, 7(4), 213 - 6, Dec. 2012 , Refereed
    Summary:The prognosis of gastric cancer patients with peritoneal metastasis is very poor. Recent findings suggest that use of trastuzumab, a monoclonal antibody-based agent that targets human epidermal growth factor receptor 2 (HER2), may improve the prognosis of gastric cancer patients with HER2 overexpression and/or gene amplification. However, whether these mechanisms of HER2 upregulation are present in gastric cancer patients with peritoneal metastasis is unclear. The status of HER2 expression in a cohort of samples obtained from 35 gastric cancer patients with peritoneal metastasis was investigated using immunohistochemistry and fluorescence in situ hybridization. In 18 cases, we also investigated the influence of induction chemotherapy on HER2 overexpression. The frequency of HER2 overexpression and gene amplification was 2.9 % (1/35) in peritoneal metastatic lesions. There was concurrence in HER2 status in the samples examined prior to and following induction of chemotherapy. Most samples from the gastric cancer patients with peritoneal metastasis did not show HER2 amplification and/or overexpression. Although our study size was small, these results suggest that trastuzumab, which is critically dependent on HER2 expression, might not be an effective agent for these patients. Consequently, other therapeutic approaches for these patients must be developed.
  • Overexpression of MDR1 and survivin, and decreased Bim expression mediate multidrug-resistance in multiple myeloma cells., Masanobu Tsubaki, Takao Satou, Tatsuki Itoh, Motohiro Imano, Makiko Komai, Minori Nishinobo, Megumi Yamashita, Masashi Yanae, Yuzuru Yamazoe, Shozo Nishida, Leukemia research, Leukemia research, 36(10), 1315 - 22, Oct. 2012 , Refereed
    Summary:Multidrug resistance represents a major obstacle for the chemotherapy of a wide variety of human tumors. To investigate the underlying mechanisms associated with resistance to anti-cancer drugs, we established anti-cancer drug-resistant multiple myeloma (MM) cell lines RPMI8226/ADM, RPMI8226/VCR, RPMI8226/DEX, and RPMI8226/L-PAM, the 50% inhibitory concentration values of which were 77-, 58-, 79-, and 30-fold higher than their parental cell lines, respectively. The resistant cell lines overexpressed MDR1 and survivin, or showed decreased Bim expression. These results indicated that regulating these factors with inhibitors might be a viable approach to increasing the susceptibility of quiescent MM cells to chemotherapy.
  • Bisphosphonate- and statin-induced enhancement of OPG expression and inhibition of CD9, M-CSF, and RANKL expressions via inhibition of the Ras/MEK/ERK pathway and activation of p38MAPK in mouse bone marrow stromal cell line ST2., Masanobu Tsubaki, Takao Satou, Tatsuki Itoh, Motohiro Imano, Masashi Yanae, Chisato Kato, Risa Takagoshi, Makiko Komai, Shozo Nishida, Molecular and cellular endocrinology, Molecular and cellular endocrinology, 361(1-2), 219 - 31, Sep. 25 2012 , Refereed
    Summary:Osteoclast differentiation is influenced by receptor activator of the NF-κB ligand (RANKL), macrophage colony-stimulating factor (M-CSF), and CD9, which are expressed on bone marrow stromal cells and osteoblasts. In addition, osteoprotegerin (OPG) is known as an osteoclastogenesis inhibitory factor. In this study, we investigated whether bisphosphonates and statins increase OPG expression and inhibit the expression of CD9, M-CSF, and RANKL in the bone marrow-derived stromal cell line ST2. We found that bisphosphonates and statins enhanced OPG mRNA expression and inhibited the expression of CD9, M-CSF, and RANKL mRNA. Futhermore, bisphosphonates and statins decreased the membrane localization of Ras and phosphorylated ERK1/2, and activated the p38MAPK. This indicates that bisphosphonates and statins enhanced OPG expression, and inhibited the expression of CD9, M-CSF, and RANKL through blocking the Ras/ERK pathway and activating p38MAPK. Accordingly, we believe that its clinical applications will be investigated in the future for the development of osteoporosis therapy.
  • A preliminary study of single intraperitoneal administration of paclitaxel followed by sequential systemic chemotherapy with S-1 plus paclitaxel for advanced gastric cancer with peritoneal metastasis., Motohiro Imano, Ying-Feng Peng, Tatsuki Itoh, Masayasu Nishikawa, Takao Satou, Atsushi Yasuda, Keisuke Inoue, Hiroaki Kato, Masayuki Shinkai, Masahiro Tsubaki, Takushi Yasuda, Haruhiko Imamoto, Shozo Nishida, Hiroshi Furukawa, Yoshifumi Takeyama, Kiyokata Okuno, Hitoshi Shiozaki, Anticancer research, Anticancer research, 32(9), 4071 - 5, Sep. 2012 , Refereed
    Summary:AIM: A preliminary study with the aim of evaluating the safety and efficacy of a single intraperitoneal administration of paclitaxel, combined with intravenous administration of paclitaxel plus S-1, was carried out in gastric cancer patients with peritoneal metastasis. PATIENTS AND METHODS: Paclitaxel was administered intraperitoneally at 80 mg/m(2). After one to two weeks, S-1 was administered at 80 mg/m(2)/day for 14 consecutive days, followed by seven days' rest. Paclitaxel was administered intravenously at 50 mg/m(2) on days 1 and 8. The safety, pharmacokinetic analysis and efficacy of this therapy were investigated. RESULTS: Fifteen patients were enrolled in this study. The toxic effects of the intraperitoneal chemotherapy were mild. The toxic effects with the systemic chemotherapy were acceptable. The ratio of (AUC peri)/(AUC pla) was 1065:1 in the pharmacokinetic analysis. The one-year overall survival rate was 10/15 (66.7%). CONCLUSION: A single intraperitoneal administration of paclitaxel combined with intravenous administration of paclitaxel plus S-1 is a well-tolerated and feasible treatment for patients with gastric cancer with peritoneal metastasis.
  • (-)-Epigallocatechin-3-gallate increases the number of neural stem cells around the damaged area after rat traumatic brain injury., Tatsuki Itoh, Motohiro Imano, Shozo Nishida, Masahiro Tsubaki, Nobuyuki Mizuguchi, Shigeo Hashimoto, Akihiko Ito, Takao Satou, Journal of neural transmission (Vienna, Austria : 1996), Journal of neural transmission (Vienna, Austria : 1996), 119(8), 877 - 90, Aug. 2012 , Refereed
    Summary:A major component of green tea is (-)-epigallocatechin gallate (EGCG), which has strong antioxidant properties. Here, we investigated the effect of EGCG on neural stem cell (NSC) proliferation around the damaged area following traumatic brain injury (TBI). In this study, male Wistar rats that had access to normal drinking water, or water containing 0.1% (w/v) EGCG, ad libitum received TBI at 10 weeks of age. Immunohistochemistry revealed that the number of nestin-positive cells around the damaged area after TBI in the EGCG treatment group increased significantly compared with the normal water group (P < 0.05). However, the number of 8-hydroxy-2'-deoxyguanosine-, 4-hydroxy-2-nonenal-, single-stranded DNA (ssDNA)-positive cells and the level of peroxidation around the damaged area after TBI significantly decreased in the EGCG treatment group when compared with the water group (P < 0.05). Furthermore, in contrast to the EGCG group, almost all ssDNA-positive cells in the water group co-localized with NeuN and nestin-staining. Ex vivo studies revealed that spheres could only be isolated from injured brain tissue in the water group at 3 days following TBI. However, in the EGCG group, spheres could be isolated at both 3 and 7 days following TBI. A greater number of spheres could be isolated from the EGCG group, which differentiated into neurons and glia in culture without basic fibroblast growth factor. These results indicate that consumption of water containing EGCG pre- and post-TBI inhibits free radical-induced degradation of NSCs, which have the potential to differentiate into neurons and glia around the area of damage following TBI.
  • [A survey of the dosage of zoledronic acid and investigation of the relationship between renal function and adverse events]., Masashi Yanae, Mariko Nakao, Kimiko Fujiwara, Akinori Kawaguchi, Masanobu Tsubaki, Yasutaka Chiba, Tetsuya Morita, Yuzuru Yamazoe, Shozo Nishida, Gan to kagaku ryoho. Cancer & chemotherapy, Gan to kagaku ryoho. Cancer & chemotherapy, 39(7), 1093 - 8, Jul. 2012 , Refereed
    Summary:Zoledronic acid(ZA)dosage should be adjusted according to the risk it poses for renal impairment. The recommended dosage for patients with creatinine clearance(Ccr)of less than 60mL/min was established on the basis of an area under the curve analysis, but is doubted because it was calculated without performing a clinical trial. Creatinine secretion from the renal tubule affects Ccr; therefore, using Ccr as the basis for dosage adjustment may be inappropriate since this can cause an overestimation of the glomerular filtration rate(GFR). The Japanese Society of Nephrology recommends using the estimated GFR(eGFR)for evaluating renal function. Therefore, this study investigated the relationship between renal function before and adverse events(AEs)after ZA administration. The dosage of only 3 of the 47 patients with Ccr less than 60mL/min could be adjusted on the basis of Ccr. During ZA therapy(3 courses), the blood urea nitrogen level and occurrence of hypokalemia were higher in the non-adjusted group than in the adjusted group, but the total number of AEs was equivalent for both groups. For all the patients, Ccr and eGFR were used as parameters for investigating AEs; the total number of AEs was equivalent for patients with differing levels of renal function. Therefore, we suggest that AEs observed during ZA therapy did not depend on the renal function level before ZA administration.
  • Reduction of metastasis, cell invasion, and adhesion in mouse osteosarcoma by YM529/ONO-5920-induced blockade of the Ras/MEK/ERK and Ras/PI3K/Akt pathway., Masanobu Tsubaki, Takao Satou, Tatsuki Itoh, Motohiro Imano, Mitsuhiko Ogaki, Masashi Yanae, Shozo Nishida, Toxicology and applied pharmacology, Toxicology and applied pharmacology, 259(3), 402 - 10, Mar. 15 2012 , Refereed
    Summary:Osteosarcoma is one of the most common primary malignant bone tumors in children and adolescents. Some patients continue to have a poor prognosis, because of the metastatic disease. YM529/ONO-5920 is a nitrogen-containing bisphosphonate that has been used for the treatment of osteoporosis. YM529/ONO-5920 has recently been reported to induce apoptosis in various tumors including osteosarcoma. However, the mode of metastasis suppression in osteosarcoma by YM529/ONO-5920 is unclear. In the present study, we investigated whether YM529/ONO-5920 inhibited tumor cell migration, invasion, adhesion, or metastasis in the LM8 mouse osteosarcoma cell line. We found that YM529/ONO-5920 significantly inhibited metastasis, cell migration, invasion, and adhesion at concentrations that did not have antiproliferative effects on LM8 cells. YM529/ONO-5920 also inhibited the mRNA expression and protein activities of matrix metalloproteinases (MMPs). In addition, YM529/ONO-5920 suppressed phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2) and the serine/threonine protein kinase B (Akt) by the inhibition of Ras prenylation. Moreover, U0126, a mitogen-activated protein kinase kinase (MEK) 1/2 inhibitor, and LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, also inhibited LM8 cell migration, invasion, adhesion, and metastasis, as well as the mRNA expression and protein activities of MMP-1, MMP-2, MMP-9, and MT1-MMP. The results indicated that YM529/ONO-5920 suppressed the Ras/MEK/ERK and Ras/PI3K/Akt pathways, thereby inhibiting LM8 cell migration, invasion, adhesion, and metastasis. These findings suggest that YM529/ONO-5920 has potential clinical applications for the treatment of tumor cell metastasis in osteosarcoma.
  • Neural stem cell proliferation surrounding the area of traumatic brain injury: Role of exercise therapy, Tatsuki Itoh, Motohiro Imano, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Akihiko Ito, Takao Satou, Stem Cells and Cancer Stem Cells, Volume 5: Therapeutic Applications in Disease and Injury, Stem Cells and Cancer Stem Cells, Volume 5: Therapeutic Applications in Disease and Injury, 59 - 72, Jan. 01 2012 , Refereed
    Summary:Exercise enhances neuronal stem cell (NSC) proliferation and neurogenesis. However, the effect of exercise on NSC proliferationsurrounding the area of damage after traumatic brain injury (TBI) isunknown. Here, we investigate the effect of running on NSCproliferation following TBI in the rat. Wistar rats received TBI andwere randomly divided into two groups: (1) non-exercise group and(2) exercise group. The exercise group ran on a treadmill for 30min/day at 22 m/min for 7 consecutive days. Immunohistochemistry wasused to monitor NSC proliferation around the damaged area and ex vivo techniques were used to isolate NSCs from the damaged region in both groups. The number of nestin-and Ki-67-positive cells observed at 3 and 7 days after TBI was significantly greater in the exercise group than in the non-exercise group (P < 0.01). Furthermore, most nestin-positive cells in the exercise group co-localized with Ki-67-positive cells. In ex vivo studies, spheres could be isolated from injured brain tissue from the exercise group at 3 and 7 days following TBI, but at only 3 days in the non-exercise group. The number of spheres isolated from injured brain tissue was greater in the exercise group than in the non-exercise group. Spheres were immunopositive for nestin and comprised of NSCs that could differentiate into neurons and glia. Exercise increases the proliferation of NSCs around the damaged area following TBI. Therefore, exercise therapy (rehabilitation) in the early phase following TBI is important for recuperation from cerebral dysfunction induced by TBI.
  • Spontaneous cerebral stroke in rats: Differentiation of new neurons from neural stem cells, Tatsuki Itoh, Kumiko Takemori, Motohiro Imano, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Hiroyuki Ito, Akihiko Ito, Takao Satou, Stem Cells and Cancer Stem Cells, Volume 2: Stem Cells and Cancer Stem Cells, Therapeutic Applications in Disease and Injury: Volume 2, Stem Cells and Cancer Stem Cells, Volume 2: Stem Cells and Cancer Stem Cells, Therapeutic Applications in Disease and Injury: Volume 2, 199 - 209, Jan. 01 2012 , Refereed
    Summary:Stroke-prone spontaneously hypertensive rats (SHRSP) are the only animal model that suffers from spontaneous cerebral stroke. In this study, we investigated the appearance of neural stem cells (NSCs) and new neurons in the penumbra and the subventricular zone (SVZ) after cerebral stroke in SHRSP. SHRSP before cerebral stroke were intraperitoneally injected with 5-bromo-2′-deoxyuridine (BrdU). SHRSP were divided into acute and chronic phase groups after cerebral stroke. Brain sections from both groups were studied with cellspecific markers such as BrdU, a cell division and proliferation marker, SOX2, a marker of NSCs, nestin, an NSC and immature astrocyte marker, doublecortin (DCX), an immature new neuron marker, and NeuN, a marker of mature neurons. NSCs and new neurons appeared in the penumbra in the early stages after cerebral stroke, and these cells differentiated into mature neurons in the chronic phase. Furthermore, soon after being affected by a cerebral stroke, there were many new neurons and immature cells, which appear to be NSCs, in the ipsilateral SVZ. The findings of the study indicate that immature cells and new neurons from the ipsilateral SVZ might migrate into the penumbra after cerebral stroke.
  • (-)-Epigallocatechin-3-gallate protects against neuronal cell death and improves cerebral function after traumatic brain injury in rats., Tatsuki Itoh, Motohiro Imano, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Akihiko Ito, Takao Satou, Neuromolecular medicine, Neuromolecular medicine, 13(4), 300 - 9, Dec. 2011 , Refereed
    Summary:A major component of green tea, a widely consumed beverage, is (-)-epigallocatechin gallate (EGCG), which has strong antioxidant properties. Our previous study has indicated that free radical production following rat traumatic brain injury (TBI) induces neural degeneration. In this study, we investigated the effects of EGCG on cerebral function and morphology following TBI. Six-week-old male Wistar rats that had access to normal drinking water, or water containing 0.1% (w/v) EGCG ad libitum, received TBI with a pneumatic controlled injury device at 10 weeks of age. Immunohistochemistry and lipid peroxidation studies revealed that at 1, 3 and 7 days post-TBI, the number of 8-hydroxy-2'-deoxyguanosine-, 4-hydroxy-2-nonenal- and single-stranded DNA (ssDNA)-positive cells, and the levels of malondialdehyde (MDA) around the damaged area after TBI, significantly decreased in the EGCG treatment group compared with the water group (P < 0.05). Most ssDNA-positive cells in the water group co-localized with neuronal cells. However, in the EGCG treatment group, few ssDNA-positive cells co-localized with neurons. In addition, there was a significant increase in the number of surviving neuronal cells and an improvement in cerebral dysfunction after TBI in the EGCG treatment group compared with the water group (P < 0.05). These results indicate that consumption of water containing EGCG pre- and post-TBI inhibits free radical-induced neuronal degeneration and apoptotic cell death around the damaged area, resulting in the improvement of cerebral function following TBI. In summary, consumption of green tea may be an effective therapy for TBI patients.
  • Exercise inhibits neuronal apoptosis and improves cerebral function following rat traumatic brain injury., Tatsuki Itoh, Motohiro Imano, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Akihiko Ito, Takao Satou, Journal of neural transmission (Vienna, Austria : 1996), Journal of neural transmission (Vienna, Austria : 1996), 118(9), 1263 - 72, Sep. 2011 , Refereed
    Summary:Exercise is reported to inhibit neuronal apoptotic cell death in the hippocampus and improve learning and memory. However, the effect of exercise on inhibition of neuronal apoptosis surrounding the area of damage after traumatic brain injury (TBI) and the improvement of cerebral dysfunction following TBI are unknown. Here, we investigate the effect of exercise on morphology and cerebral function following TBI in rats. Wistar rats received TBI by a pneumatic controlled injury device were randomly divided into two groups: (1) non-exercise group and (2) exercise group. The exercise group ran on a treadmill for 30 min/day at 22 m/min for seven consecutive days. Immunohistochemical and behavioral studies were performed following TBI. The number of single-stranded DNA (ssDNA)-positive cells around the damaged area early after TBI was significantly reduced in the exercise group compared with the non-exercise group (P < 0.05). Furthermore, most ssDNA-positive cells in the non-exercise group co-localized with neuronal cells. However, in the exercise group, a few ssDNA-positive cells co-localized with neurons. In addition, there was a significant increase in neuronal cell number and improvement in cerebral dysfunction after TBI in the exercise group compared with the non-exercise group (P < 0.05). These results indicate that exercise following TBI inhibits neuronal degeneration and apoptotic cell death around the damaged area, which results in improvement of cerebral dysfunction. In summary, treadmill running improved cerebral dysfunction following TBI, indicating its potential as an effective clinical therapy. Therefore, exercise therapy (rehabilitation) in the early phase following TBI is important for recuperation from cerebral dysfunction.
  • Statin-induced apoptosis via the suppression of ERK1/2 and Akt activation by inhibition of the geranylgeranyl-pyrophosphate biosynthesis in glioblastoma., Masashi Yanae, Masanobu Tsubaki, Takao Satou, Tatsuki Itoh, Motohiro Imano, Yuzuru Yamazoe, Shozo Nishida, Journal of experimental & clinical cancer research : CR, Journal of experimental & clinical cancer research : CR, 30, 74 - 74, Aug. 10 2011 , Refereed
    Summary:BACKGROUND: Statins are inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, the rate-limiting enzyme in cholesterol synthesis. The inhibition of this key enzyme in the mevalonate pathway leads to suppression of cell proliferation and induction of apoptosis. However, the molecular mechanism of apoptosis induction by statins is not well understood in glioblastoma. In the present study, we attempted to elucidate the mechanism by which statins induce apoptosis in C6 glioma cells. METHODS: The cytotoxicity of statins toward the C6 glioma cells were evaluated using a cell viability assay. The enzyme activity of caspase-3 was determined using activity assay kits. The effects of statins on signal transduction molecules were determined by western blot analyses. RESULTS: We found that statins inhibited cell proliferation and induced apoptosis in these cells. We also observed an increase in caspase-3 activity. The apoptosis induced by statins was not inhibited by the addition of farnesyl pyrophosphate, squalene, ubiquinone, and isopentenyladenine, but by geranylgeranyl-pyrophosphate (GGPP). Furthermore, statins decreased the levels of phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt. CONCLUSIONS: These results suggest that statins induce apoptosis when GGPP biosynthesis is inhibited and consequently decreases the level of phosphorylated ERK1/2 and Akt. The results of this study also indicate that statins could be used as anticancer agents in glioblastoma.
  • [Clinical evaluation of calculating carboplatin dosage using Japanese equation for estimating GFR for gynecologic cancer]., Yasuhiro Kidera, Masahiko Nakao, Masanobu Tsubaki, Masahiro Yoshinaga, Fumihiro Kajitani, Masashi Yanae, Mika Sakano, Yuzuru Yamazoe, Yasutaka Chiba, Kenzo Moriyama, Shozo Nishida, Gan to kagaku ryoho. Cancer & chemotherapy, Gan to kagaku ryoho. Cancer & chemotherapy, 38(7), 1143 - 8, Jul. 2011 , Refereed
    Summary:Glomerular filtration rate (GFR) is an important factor when considering carboplatin dosage adjustment. The Japanese equation for estimating GFR (eGFR) was recommended as a guideline for evaluating GFR in 2009 by The Japanese Society of Nephrology. However, benefits in the field of cancer chemotherapy with the use of eGFR have not yet been shown. To clarify the clinical benefits of eGFR, we investigated the renal function of 100 patients with gynecologic cancer who were treated with carboplatin from 2003 through 2009, and the carboplatin dosage was calculated by the Calvert formula in which eGFR was substituted for GFR. To predict the clinical benefit on the basis of carboplatin dosage using eGFR, we retrospectively divided the patients into two groups so that carboplatin dosage was within dosage in using eGFR and one was not. We compared response rates and adverse effects of the two groups. Renal function using eGFR was lower than renal function calculated by using the other formulae. Carboplatin dosage using eGFR was significantly lower than the dosage calculated with the other formulae (p<0.01). Moreover, the patients group actually, administered the dosage calculated by eGFR showed less side effects than the group of patients not treated this way, but the efficacy did not change. Thus, using eGFR in planning carboplatin dosage suggested clinical application to patients with Japanese gynecologic cancer.
  • High-dose dexamethasone plus antihistamine prevents colorectal cancer patients treated with modified FOLFOX6 from hypersensitivity reactions induced by oxaliplatin., Yasuhiro Kidera, Taroh Satoh, Shinya Ueda, Wataru Okamoto, Isamu Okamoto, Soichi Fumita, Kimio Yonesaka, Hidetoshi Hayashi, Chihiro Makimura, Kunio Okamoto, Hidemi Kiyota, Junji Tsurutani, Masaki Miyazaki, Masahiro Yoshinaga, Kimiko Fujiwara, Yuzuru Yamazoe, Kenzo Moriyama, Masanobu Tsubaki, Yasutaka Chiba, Shozo Nishida, Kazuhiko Nakagawa, International journal of clinical oncology, International journal of clinical oncology, 16(3), 244 - 9, Jun. 2011 , Refereed
    Summary:BACKGROUND: Oxaliplatin is a third-generation platinum compound and a key agent for the management of colorectal cancer. Patients treated with oxaliplatin are at risk for hypersensitivity reactions. We designed a modified premedication regimen to prevent oxaliplatin-related hypersensitivity reactions and assessed if this approach is effective. METHODS: A retrospective cohort study of patients with advanced colorectal cancer who received modified FOLFOX6 (mFOLFOX6) was performed. Patients received routine premedication with dexamethasone 8 mg and granisetron 3 mg for the first five cycles of mFOLFOX6. From the sixth cycle onward, cohort 1 received the same premedication, and cohort 2 received modified premedication (diphenhydramine 50 mg orally, followed by dexamethasone 20 mg, granisetron 3 mg, and famotidine 20 mg). We compared the incidence of hypersensitivity reactions, duration of treatment, and reasons for treatment withdrawal between the two cohorts. RESULTS: A total of 181 patients were studied (cohort 1, 81; cohort 2, 100). Hypersensitivity reactions developed in 16 patients (20%) in cohort 1 and 7 (7.0%) in cohort 2 (P = 0.0153). The median number of cycles increased from 9 in cohort 1 to 12 in cohort 2. Apart from progressive disease, neurotoxicity was the reason for discontinuing treatment in 20% of the patients in cohort 1, as compared with 53% in cohort 2. CONCLUSION: Increased doses of dexamethasone and antihistamine significantly reduced oxaliplatin-related hypersensitivity reactions. This effective approach should be considered for all patients who receive FOLFOX, allowing treatment to be completed as planned.
  • Association between occupational exposure levels of antineoplastic drugs and work environment in five hospitals in Japan., Jin Yoshida, Shigeki Koda, Shozo Nishida, Toshiaki Yoshida, Keiko Miyajima, Shinji Kumagai, Journal of oncology pharmacy practice : official publication of the International Society of Oncology Pharmacy Practitioners, Journal of oncology pharmacy practice : official publication of the International Society of Oncology Pharmacy Practitioners, 17(1), 29 - 38, Mar. 2011 , Refereed
    Summary:PURPOSE: The aim of the present study was to evaluate the measurement of contamination by antineoplastic drugs for safer handling of such drugs by medical workers. We investigated the relationship between the contamination level of antineoplastic drugs and the conditions of their handling. METHODS: Air samples and wipe samples were collected from equipment in the preparation rooms of five hospitals (hospitals A-E). These samples were subjected to measurement of the amounts of cyclophosphamide (CPA), fluorouracil (5FU), gemcitabine (GEM), and platinum-containing drugs (Pt). Twenty-four-hour urine samples were collected from the pharmacists who handled or audited, the antineoplastic drugs were analyzed for CPA and Pt. RESULTS: Pt was detected from air samples inside BSC in hospital B. Antineoplastic drugs were detected from wipe samples of the BSC in hospitals A, B, D, and E and of other equipment in the preparation rooms in hospitals A, B, C, and D. Cyclophosphamide and 5FU were detected from wipe samples of the air-conditioner filter in hospital A, and CPA was detected from that in hospital D. Cyclophosphamide was detected from urine samples of workers in hospitals B, D, and E. CONCLUSION: The contamination level of antineoplastic drugs was suggested to be related with the amount of drugs handled, cleaning methods of the equipment, and the skill level of the technique of maintaining negative pressure inside a vial. In order to reduce the contamination and exposure to antineoplastic drugs in the hospital work environment very close to zero, comprehensive safety precautions, including adequate mixing and cleaning methods was required in addition to BSC and closed system device.
  • Exercise increases neural stem cell proliferation surrounding the area of damage following rat traumatic brain injury., Tatsuki Itoh, Motohiro Imano, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Akihiko Ito, Takao Satou, Journal of neural transmission (Vienna, Austria : 1996), Journal of neural transmission (Vienna, Austria : 1996), 118(2), 193 - 202, Feb. 2011 , Refereed
    Summary:Exercise enhances neuronal stem cell (NSC) proliferation and neurogenesis. However, the effect of exercise on NSC proliferation surrounding the area of damage after traumatic brain injury (TBI) is unknown. Here, we investigate the effect of running on NSC proliferation following TBI in the rat. Wistar rats received TBI and were randomly divided into two groups: (1) non-exercise group and (2) exercise group. The exercise group ran on a treadmill for 30 min/day at 22 m/min for 7 consecutive days. Immunohistochemistry was used to monitor NSC proliferation around the damaged area, and ex vivo techniques were used to isolate NSCs from the damaged region in both groups. The number of nestin- and Ki67-positive cells observed at 3 and 7 days after TBI was significantly greater in the exercise group than in the non-exercise group (P < 0.01). Furthermore, most nestin-positive cells in the exercise group co-localized with Ki67-positive cells. In ex vivo studies, spheres could be isolated from injured brain tissue from the exercise group at 3 and 7 days following TBI, but at only 3 days in the non-exercise group. The number of spheres isolated from injured brain tissue was greater in the exercise group than in the non-exercise group. Spheres were immunopositive for nestin and comprised NSCs that could differentiate into neurons and glia. Exercise increases the proliferation of NSCs around the damaged area following TBI. Therefore, exercise therapy (rehabilitation) in the early phase following TBI is important for recuperation from cerebral dysfunction induced by TBI.
  • Impact of Intraperitoneal Chemotherapy after Gastrectomy with Positive Cytological Findings in Peritoneal Washings, M. Imano, H. Imamoto, T. Itoh, T. Satou, Y. F. Peng, A. Yasuda, H. Kato, K. Nishiki, O. Shiraishi, M. Shinkai, M. Tsubaki, T. Yasuda, S. Nishida, Y. Takeyama, K. Okuno, H. Shiozaki, EUROPEAN SURGICAL RESEARCH, EUROPEAN SURGICAL RESEARCH, 47(4), 254 - 259, 2011 , Refereed
    Summary:Background: There is no standard treatment available for gastric cancer patients whose sole 'non-curative factor' is positive cytological findings in peritoneal washings (CFPW). The aim of this study was to examine the safety, pharmacokinetics and efficacy for free intraperitoneal cancer cells of intraperitoneal chemotherapy with paclitaxel after gastrectomy with en bloc D2 lymph node dissection in cases of gastric cancer with positive CFPW. Methods: Ten patients with gastric cancer who underwent gastrectomy and systemic lymphadenectomy with D2 dissection, without any other non-curative factors besides positive CFPW, were treated with early postoperative intraperitoneal paclitaxel. Intra-chemotherapeutic toxicity and operative complications were measured using NCI-CTC version 3.0. Intraperitoneal and plasma paclitaxel concentrations were measured using a high-performance liquid chromatographic assay. Results: Grade 3/4 toxic effects included anemia (20%) and neutropenia (10%) that required no treatment. Operative complications were, for example, superficial surgical site infections (10%) that were treated with antibiotics. No viable cancer cells were observed in the intra-abdominal fluid 24 h after intraperitoneal administration of paclitaxel. The intraperitoneal/plasma area under the drug concentration-time curve ratio was 2,003.3: 1. Conclusion: Intraperitoneal chemotherapy with paclitaxel is a safe and effective treatment modality for free intraperitoneal cancer cells. Copyright (C) 2011 S. Karger AG, Basel
  • 含窒素ビスホスホネートYM529/ONO-5920はRho/ROCK経路抑制を介してマウス悪性黒色腫の腫瘍転移を阻害する(Nitrogen-containing bisphosphonate, YM529/ONO-5920, inhibits tumor metastasis in mouse melanoma through suppression of the Rho/ROCK pathway), 椿 正寛, 金子 淳一, 木寺 康裕, 谷森 佳弘, 柳江 正嗣, 西田 升三, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回(7), 4P - 0990, Dec. 2010
    Summary:The small GTPases of the Ras and Rho families are widely involved in tumorigenesis and metastasis. We recently showed that YM529/ONO-5920, a new developed bisphosphonate, inhibits the mevalonate pathway, is required for the prenylation of the small GTPases. In this study, we investigated whether YM529/ONO-5920 inhibits tumor cell migration, invasion, adhesion, and metastasis in B16BL6 cells, a mouse melanoma cell line. It was found that YM529/ONO-5920 significantly inhibited lung metastasis, cell migration, invasion, and adhesion at concentrations that did not show anti-proliferative effects on B16BL6 cells. YM529/ONO-5920 also inhibited the expression of matrix metalloproteinases (MMPs) and very late antigens (VLAs). Furthermore, YM529/ONO-5920 suppressed Rho activation, but not activation of Ras. The results indicate that YM529/ONO-5920 suppresses the Rho/ROCK pathways, thereby inhibiting B16BL6 cell migration, invasion, adhesion and metastasis. These findings suggest that YM529/ONO-5920 has potential clinical applications for the treatment of tumour cell metastasis.
  • Reduction of lung metastasis, cell invasion, and adhesion in mouse melanoma by statin-induced blockade of the Rho/Rho-associated coiled-coil-containing protein kinase pathway., Yasuhiro Kidera, Masanobu Tsubaki, Yuzuru Yamazoe, Kaori Shoji, Haruyuki Nakamura, Mitsuhiko Ogaki, Takao Satou, Tatsuki Itoh, Misako Isozaki, Junichi Kaneko, Yoshihiro Tanimori, Masashi Yanae, Shozo Nishida, Journal of experimental & clinical cancer research : CR, Journal of experimental & clinical cancer research : CR, 29(1), 127 - 127, Sep. 16 2010 , Refereed
    Summary:BACKGROUND: Melanomas are highly malignant and have high metastatic potential; hence, there is a need for new therapeutic strategies to prevent cell metastasis. In the present study, we investigated whether statins inhibit tumor cell migration, invasion, adhesion, and metastasis in the B16BL6 mouse melanoma cell line. METHODS: The cytotoxicity of statins toward the B16BL6 cells were evaluated using a cell viability assay. As an experimental model, B16BL6 cells were intravenously injected into C57BL/6 mice. Cell migration and invasion were assessed using Boyden chamber assays. Cell adhesion analysis was performed using type I collagen-, type IV collagen-, fibronectin-, and laminin-coated plates. The mRNA levels, enzyme activities and protein levels of matrix metalloproteinases (MMPs) were determined using RT-PCR, activity assay kits, and Western blot analysis, respectively; the mRNA and protein levels of vary late antigens (VLAs) were also determined. The effects of statins on signal transduction molecules were determined by western blot analyses. RESULTS: We found that statins significantly inhibited lung metastasis, cell migration, invasion, and adhesion at concentrations that did not have cytotoxic effects on B16BL6 cells. Statins also inhibited the mRNA expressions and enzymatic activities of matrix metalloproteinases (MMPs). Moreover, they suppressed the mRNA and protein expressions of integrin α2, integrin α4, and integrin α5 and decreased the membrane localization of Rho, and phosphorylated LIM kinase (LIMK) and myosin light chain (MLC). CONCLUSIONS: The results indicated that statins suppressed the Rho/Rho-associated coiled-coil-containing protein kinase (ROCK) pathways, thereby inhibiting B16BL6 cell migration, invasion, adhesion, and metastasis. Furthermore, they markedly inhibited clinically evident metastasis. Thus, these findings suggest that statins have potential clinical applications for the treatment of tumor cell metastasis.
  • Edaravone protects against apoptotic neuronal cell death and improves cerebral function after traumatic brain injury in rats., Tatsuki Itoh, Takao Satou, Shozo Nishida, Masahiro Tsubaki, Motohiro Imano, Shigeo Hashimoto, Hiroyuki Ito, Neurochemical research, Neurochemical research, 35(2), 348 - 55, Feb. 2010 , Refereed
    Summary:Edaravone is a novel free radical scavenger used clinically in patients with acute cerebral infarction; however, it has not been assessed in traumatic brain injury (TBI). We investigated the effects of edaravone on cerebral function and morphology following TBI. Rats received TBI with a pneumatic controlled injury device. Edaravone (3 mg/kg) or physiological saline was administered intravenously following TBI. Numbers of 8-OHdG-, 4-HNE-, and ssDNA-positive cells around the damaged area after TBI were significantly decreased in the edaravone group compared with the saline group (P < 0.01). There was a significant increase in neuronal cell number and improvement in cerebral dysfunction after TBI in the edaravone group compared with the saline group (P < 0.01). Edaravone administration following TBI inhibited free radical-induced neuronal degeneration and apoptotic cell death around the damaged area. In summary, edaravone treatment improved cerebral dysfunction following TBI, suggesting its potential as an effective clinical therapy.
  • The novel free radical scavenger, edaravone, increases neural stem cell number around the area of damage following rat traumatic brain injury., Tatsuki Itoh, Takao Satou, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Hiroyuki Ito, Neurotoxicity research, Neurotoxicity research, 16(4), 378 - 89, Nov. 2009 , Refereed
    Summary:Edaravone is a novel free radical scavenger that is clinically employed in patients with acute cerebral infarction, but has not previously been used to treat traumatic brain injury (TBI). In this study, we investigated the effect of edaravone administration on rat TBI. In particular, we used immunohistochemistry to monitor neural stem cell (NSC) proliferation around the area damaged by TBI. Two separate groups of rats were administered saline or edaravone (3 mg/kg) after TBI and then killed chronologically. We also used ex vivo techniques to isolate NSCs from the damaged region and observed nestin-positive cells at 1, 3, and 7 days following TBI in both saline- and edaravone-treated groups. At 3 days following TBI in both groups, there were many large cells that morphologically resembled astrocytes. At 1 and 7 days following TBI in the saline group, there were a few small nestin-positive cells. However, in the edaravone group, there were many large nestin-positive cells at 7 days following TBI. At 3 and 7 days following TBI, the number of nestin-positive cells in the edaravone group increased significantly compared with the saline group. There were many single-stranded DNA-, 8-hydroxy-2'-deoxyguanosine-, and 4-hydroxy-2-nonenal-positive cells in the saline group following TBI, but only a few such cells in the edaravone group following TBI. Furthermore, almost all ssDNA-positive cells in the saline group co-localized with Hu, nestin, and glial fibrillary acidic protein (GFAP) staining, but not in the edaravone group. In the ex vivo study, spheres could only be isolated from injured brain tissue in the saline group at 3 days following TBI. However, in the edaravone group, spheres could be isolated from injured brain tissue at both 3 and 7 days following TBI. The number of spheres isolated from injured brain tissue in the edaravone group showed a significant increase compared with the saline group. The spheres isolated from both saline and edaravone groups were immunopositive for nestin, but not Tuj1 or vimentin. Moreover, the spheres differentiated into Tuj1-, GFAP-, and O4-positive cells after 4 days in culture without bFGF. This result indicated that the spheres were neurospheres composed of NSCs that could differentiate into neurons and glia. Edaravone administration inhibited production of free radicals known to induce neuronal degeneration and cell death after brain injury, and protected nestin-positive cells, including NSCs, with the potential to differentiate into neurons and glia around the area damaged by TBI.
  • Dimethylfumarate inhibits tumor cell invasion and metastasis by suppressing the expression and activities of matrix metalloproteinases in melanoma cells., Yuzuru Yamazoe, Masanobu Tsubaki, Hiroshi Matsuoka, Takao Satou, Tatsuki Itoh, Takashi Kusunoki, Yasuhiro Kidera, Yoshihiro Tanimori, Kaori Shoji, Haruyuki Nakamura, Mitsuhiko Ogaki, Saori Nishiura, Shozo Nishida, Cell biology international, Cell biology international, 33(10), 1087 - 94, Oct. 2009 , Refereed
    Summary:NF-kappaB acts as a signal transducer during tumor progression, cell invasion, and metastasis. Dimethylfumarate (DMF) is reported to inhibit tumor necrosis factor-alpha-induced nuclear entry of NF-kappaB/p65. However, only a few reports suggest that DMF inhibits tumor metastasis; also the molecular mechanisms underlying the inhibition of metastasis are poorly understood. We investigated the inhibition of tumor invasion and metastasis by DMF in a melanoma cell line, B16BL6. DMF inhibited B16BL6 cell invasion and metastasis by suppressing the expression and activities of MMPs. DMF also inhibited the nuclear entry of NF-kappaB/p65, thus inhibiting B16BL6 cell invasion and metastasis. These results suggest that DMF is potentially useful as an anti-metastatic agent for the treatment of malignant melanoma.
  • Tamoxifen inhibits tumor cell invasion and metastasis in mouse melanoma through suppression of PKC/MEK/ERK and PKC/PI3K/Akt pathways., Hiroshi Matsuoka, Masanobu Tsubaki, Yuzuru Yamazoe, Mitsuhiko Ogaki, Takao Satou, Tatsuki Itoh, Takashi Kusunoki, Shozo Nishida, Experimental cell research, Experimental cell research, 315(12), 2022 - 32, Jul. 15 2009 , Refereed
    Summary:In melanoma, several signaling pathways are constitutively activated. Among these, the protein kinase C (PKC) signaling pathways are activated through multiple signal transduction molecules and appear to play major roles in melanoma progression. Recently, it has been reported that tamoxifen, an anti-estrogen reagent, inhibits PKC signaling in estrogen-negative and estrogen-independent cancer cell lines. Thus, we investigated whether tamoxifen inhibited tumor cell invasion and metastasis in mouse melanoma cell line B16BL6. Tamoxifen significantly inhibited lung metastasis, cell migration, and invasion at concentrations that did not show anti-proliferative effects on B16BL6 cells. Tamoxifen also inhibited the mRNA expressions and protein activities of matrix metalloproteinases (MMPs). Furthermore, tamoxifen suppressed phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt through the inhibition of PKCalpha and PKCdelta phosphorylation. However, other signal transduction factor, such as p38 mitogen-activated protein kinase (p38MAPK) was unaffected. The results indicate that tamoxifen suppresses the PKC/mitogen-activated protein kinase kinase (MEK)/ERK and PKC/phosphatidylinositol-3 kinase (PI3K)/Akt pathways, thereby inhibiting B16BL6 cell migration, invasion, and metastasis. Moreover, tamoxifen markedly inhibited not only developing but also clinically evident metastasis. These findings suggest that tamoxifen has potential clinical applications for the treatment of tumor cell metastasis.
  • Improvement of cerebral function by anti-amyloid precursor protein antibody infusion after traumatic brain injury in rats., Tatsuki Itoh, Takao Satou, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Hiroyuki Ito, Molecular and cellular biochemistry, Molecular and cellular biochemistry, 324(1-2), 191 - 9, Apr. 2009 , Refereed
    Summary:We previously demonstrated the increased amyloid precursor protein (APP) immunoreactivity around the site of damage after traumatic brain injury (TBI). However, the function of APP after TBI has not been evaluated. In this study, we investigated the effects of direct infusion of an anti-APP antibody into the damaged brain region on cerebral function and morphological changes following TBI in rats. Three days after TBI, there were many TUNEL-positive neurons and astrocytes around the damaged region and a significantly greater number of TUNEL-positive cells in the PBS group compared with the anti-APP group found. Seven days after TBI, there were significantly a greater number of large glial fibrillary acidic protein-positive cells, long elongated projections, and microtubule-associated protein-2-positive cells around the damaged region in the anti-APP group compared with the PBS group found. Seven days after TBI, the region of brain damage was significantly smaller and the time to arrival at a platform was significantly shorter in the anti-APP group compared with the PBS group. Furthermore, after TBI in the anti-APP group, the time to arrival at the platform recovered to that observed in uninjured sham operation group rats. These data suggest that the overproduction of APP after TBI inhibits astrocyte activity and reduces neural cell survival around the damaged brain region, which speculatively may be related to the induction of Alzheimer disease-type dementia after TBI.
  • The relationship between SDF-1alpha/CXCR4 and neural stem cells appearing in damaged area after traumatic brain injury in rats., Tatsuki Itoh, Takao Satou, Hiroyuki Ishida, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Hiroyuki Ito, Neurological research, Neurological research, 31(1), 90 - 102, Feb. 2009 , Refereed
    Summary:OBJECTIVE: The actual relationship between neural stem cells and SDF-1alpha/CXCR4 after brain injury has not yet been elucidated, although recent studies have speculated that stromal cell-derived factor-1alpha (SDF-1alpha) and its receptor, CXCR4, could contribute to neural stem cells migration after brain injury. In the present study, the temporal relationship between neural stem cells (NSCs) and SDF-1alpha/CXCR4 around a damaged area was investigated using a rat traumatic brain injury (TBI) model. METHODS: We used molecular biology techniques and immunohistochemistry to investigate the relationship between SDF-1alpha/CXCR4 expression and NSCs existence around a damaged area after TBI in the rat brain. RESULTS: SDF-1alpha mRNA expression and SDF-1alpha protein synthesis did not increase after TBI. However, SDF-1alpha leaked from the injured area and diffused into the cortex 1-3 days after TBI. Subsequently, the levels of CXCR4 mRNA expression and CXCR4 protein synthesis increased significantly. Many small cells with a nestin-positive cytoplasm and fibers also showed immunopositivity for both CXCR4 and SOX-2, but not for GFAP, 3-7 days after TBI. Moreover, a proportion of the CXCR4-positive cells and fibers also showed immunostaining for neurofilaments. DISCUSSION: These results suggest that the leaked SDF-1alpha attracted CXCR4-positive NSCs as well as elongated nerve fibers. It is considered that the SDF-1alpha/CXCR4 system in the brain contributes to neural stem cells appearance and maturation after TBI. Therefore, exploitation of the SDF-1alpha/CXCR4 system around a damaged area may improve the brain dysfunction after TBI.
  • Expression of amyloid precursor protein after rat traumatic brain injury., Tatsuki Itoh, Takao Satou, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Hiroyuki Ito, Neurological research, Neurological research, 31(1), 103 - 9, Feb. 2009 , Refereed
    Summary:OBJECTIVE: Previous reports have demonstrated that some focal brain injuries increase amyloid precursor protein (APP) immunoreactivity in the region surrounding the injury in the cerebral cortex. However, the chronologic changes in APP expression have not been evaluated after traumatic brain injury (TBI). METHODS: In this study, we immunohistochemically and biologically investigated chronologic changes in cellular sources and levels of APP production after rat TBI. RESULTS: In the present report, we show that traumatic brain injury increased the expression of APP in the neuronal perikarya and in damaged dystrophic neurites from 1 to 90 days after injury. Moreover, 7 days after injury, some macrophages/microglia also were co-localized with APP, which was overproduced by the neuronal perikarya and APP-positive dystrophic neurites after injury and then APP were phagocytosed by macrophages/microglia during this phase. However, astroglia did not express APP immunopositivity after brain injury. DISCUSSION: These results suggested that long-term overexpression of APP was confirmed by immunohistochemical and biologic technique after TBI. This may be related to the induction of Alzheimer type dementia and it is a very important risk factor for this disease.
  • Neuroprotective effects of an extract from the inflamed skin of rabbits inoculated with vaccinia virus on glutamate-induced neurotoxicity in cultured neuronal cell line., Tatsuki Itoh, Takao Satou, Shozo Nishida, Masahiro Tsubaki, Shigeo Hashimoto, Hiroyuki Ito, Neurological research, Neurological research, 30(4), 430 - 4, May 2008 , Refereed
    Summary:OBJECTIVE: Protein-free extracts from the inflamed skin of rabbits inoculated with vaccinia virus (Rosemorgen and Neurotropin are widely employed to combat chronic pain and treat allergic conditions in human subjects in Japan. However, the pharmacologic mechanisms of Rosemorgen and Neurotropin remain unclear. METHODS: In this study, we examined the effects of Rosemorgen on L-glutamic acid (Glu)-induced cell death in N18-RE-105 neural cell line, which only possessed non-N-methyl-D-aspartate (NMDA)-type receptors. RESULTS: There were many large cytoplasmic cells and elongation of fivers in phosphate-buffered saline (PBS) additional group without Glu. In PBS and Glu simultaneous additional group, the survival ratio was decrease significantly compared with PBS alone group. Moreover, there were dead cells which did not have cytoplasm and aggregated nucleus. The Glu-induced cell death of N18-RE-105 cells was inhibited by both pre-treatment (24 hours before Glu treatment) and simultaneous treatment with Rosemorgen. There were many large cytoplasmic cells and elongation of fivers in Rosemorgen group. DISCUSSION: From this finding in N18-RE-105 cells, Rosemorgen was concluded to inhibit Glu-induced cell death via non-NMDA type receptors. One of the pharmacologic mechanisms of Rosemorgen has been clear. These results suggest that Rosemorgen depresses allodynia and chronic pain through interaction with non-NMDA type receptors.
  • [Fluvastatin induces apoptosis on human tongue carcinoma cell line HSC-3]., Kimiko Fujiwara, Masanobu Tsubaki, Yuzuru Yamazoe, Saori Nishiura, Takeru Kawaguchi, Mitsuhiko Ogaki, Minori Nishinobo, Kenji Shimamoto, Kenzo Moriyama, Shozo Nishida, Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan, Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan, 128(1), 153 - 8, Jan. 2008 , Refereed
    Summary:Statins, which are inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, suppress cell proliferation and induce apoptosis in various cancer cell lines. However, the effects of statins in head and neck carcinoma have not been reported. In this study, we investigated the mechanism by which fluvastatin induces apoptosis in HSC-3 cells. An increase in caspase-3 activity was observed. The apoptosis induced by fluvastatin was inhibited by the addition of geranylgeranyl pyrophosphate (GGPP) to the cell culture. When we examined the survival signals at the time of apoptotic induction, we also found that fluvastatin had caused a remarkable decrease in the phosphorylation of extracellular signal-regulated kinase (ERK) 1/2. Moreover, we also found that U0126, a MEK1/2 inhibitor, induces apoptosis in HSC-3 cells. These results suggested that fluvastatin induces apoptosis by inhibiting GGPP biosynthesis and consequently decreasing the level of phosphorylated ERK1/2. The results of this study also indicate that fluvastatin may be used as an anticancer agent for tongue carcinoma.
  • Nitrogen-containing bisphosphonate, YM529/ONO-5920, inhibits macrophage inflammatory protein 1 alpha expression and secretion in mouse myeloma cells., Masanobu Tsubaki, Chisato Kato, Minori Nishinobo, Mitsuhiko Ogaki, Takao Satou, Tatsuki Ito, Takashi Kusunoki, Kimiko Fujiwara, Yuzuru Yamazoe, Shozo Nishida, Cancer science, Cancer science, 99(1), 152 - 8, Jan. 2008 , Refereed
    Summary:Macrophage inflammatory protein 1 alpha (MIP-1 alpha) is detected at high concentrations in patients with multiple myeloma, and it is thought to play an important role in the etiology of multiple myeloma and osteolysis. Thus, we investigated whether or not YM529/ONO-5920, a new bisphosphonate, inhibited MIP-1 alpha mRNA expression in, and MIP-1 alpha secretion from, mouse myeloma cells. When the cells were stimulated by lipopolysaccharide, increased MIP-1 alpha mRNA expression and MIP-1 alpha secretion were observed. YM529/ONO-5920 inhibited MIP-1 alpha mRNA expression and MIP-1 alpha secretion in a concentration-dependent manner. A transient increase in the phosphorylation of extracellular-regulated kinase 1/2 (ERK1/2) and Akt was observed after lipopolysaccharide stimulation. After YM529/ONO-5920 was given, there was no transient increase in the phosphorylation of ERK1/2 or Akt. These results indicated that YM529/ONO-5920 inhibited the expression and secretion of MIP-1 alpha through blocking the signaling pathway of the Ras/mitogen-activated protein kinase kinase/ERK and Ras/phosphatidylinositol-3 kinase/Akt. Accordingly, YM529/ONO-5920 appears to have promise for use in effective future therapy for osteolysis and myeloma cell growth that depends on MIP-1 alpha.
  • Actual conditions of the mixing of antineoplastic drugs for injection in hospitals in Osaka Prefecture, Japan., Jin Yoshida, Hiroshi Kosaka, Shozo Nishida, Shinji Kumagai, Journal of occupational health, Journal of occupational health, 50(1), 86 - 91, 2008 , Refereed
    Summary:We conducted a questionnaire survey in order to grasp the actual conditions under which antineoplastic drugs are mixed for injection in hospitals. Questionnaires were sent to all 155 hospitals with 100 or more beds for general patients in Osaka Prefecture, Japan. The response rate was 69.0%. Mixing of antineoplastic drugs was done in 81.3% of the hospitals. The questionnaire was answered by doctors in 17.2% of the hospitals with antineoplastic drugs, nurses in 11.5%, and pharmacists in 70.1%. Mixing of antineoplastic drugs was done by doctors in 58.6% of the hospitals, nurses in 44.8%, and pharmacists in 63.2% (multiple answers). Occupational exposure to antineoplastic drugs was recognized in 97.7% of the hospitals. The mean frequency of the mixing operation was 8.8 d per month per worker. The mean number of antineoplastic drugs handled was 7.4 types. Guidelines for the safe handling of antineoplastic drugs were used in 52.8% of the hospitals and a biological safety cabinet was available in 57.4%. Gloves, mask, gown and goggles were used in 82.7, 69.0, 62.1 and 36.8% of the hospitals, respectively, but no personal protective equipment was used in 10.1%. The safety precautions of the hospitals in which the number of beds was small tended to be fewer than those of the hospitals in which the number of beds was large. Used vials and ampoules were disposed of as clinical, infectious or exclusive antineoplastic drug waste by 74.7% of the hospitals. Safety measures for handling the excrement of patients treated with antineoplastic drugs were performed in 8.0% of the hospitals. In 43.7% of the hospitals, the responders had experienced accidents during antineoplastic drug preparation, such as drugs adhering to hands or eyes, drug leakage, accidental injection and cutting by ampoules. Because of the adverse effects of antineoplastic drugs, all hospitals in which the healthcare workers handle them should promote safety precautions.
  • Extramedullary plasmacytoma of the larynx: a case report from Japan., Takeshi Kusunoki, Katsuhisa Ikeda, Kiyotaka Murata, Shozo Nishida, Masanobu Tsubaki, Ear, nose, & throat journal, Ear, nose, & throat journal, 86(12), 763 - 4, Dec. 2007 , Refereed
    Summary:Laryngeal plasmacytoma is rare in Japan; to the best of our knowledge, only 8 other cases have been previously reported. We report a new case of extramedullary plasmacytoma of the larynx in a 76-year-old Japanese woman. Immunohistochemical investigation revealed that her disease had not progressed to multiple myeloma. The patient declined radiotherapy, and her condition remained stable at the 6-month follow-up.
  • Macrophage inflammatory protein-1alpha (MIP-1alpha) enhances a receptor activator of nuclear factor kappaB ligand (RANKL) expression in mouse bone marrow stromal cells and osteoblasts through MAPK and PI3K/Akt pathways., Masanobu Tsubaki, Chisato Kato, Miyuki Manno, Mitsuhiko Ogaki, Takao Satou, Tatsuki Itoh, Takashi Kusunoki, Yoshihiro Tanimori, Kimiko Fujiwara, Hiroshi Matsuoka, Shozo Nishida, Molecular and cellular biochemistry, Molecular and cellular biochemistry, 304(1-2), 53 - 60, Oct. 2007 , Refereed
    Summary:Osteolytic lesions are rapidly progressive during the terminal stages of myeloma, and the bone pain or bone fracture that occurs at these lesions decreases the patients' quality of life to a notable degree. In relation to the etiology of this bone destruction, it has been reported recently that MIP-1alpha, produced in large amounts in myeloma patients, acts indirectly on osteoclastic precursor cells, and activates osteoclasts by way of bone-marrow stromal cells or osteoblasts, although the details of this process remain obscure. In the present study, our group investigated the mechanism by which RANKL expression is induced by MIP-1alpha and the effects of MIP-1alpha on the activation of osteoclasts. RANKL mRNA and RANKL protein expressions increased in both ST2 cells and MC3T3-E1 cells in a MIP-1alpha concentration-dependent manner. RANKL mRNA expression began to increase at 1 h after the addition of MIP-1alpha; the increase became remarkable at 2 h, and continuous expression was observed subsequently. Both ST2 and MC3T3-E1 cells showed similar levels of increased RANKL protein expression at 1, 2, and 3 days after the addition of MIP-1alpha. After the addition of MIP-1alpha, the amount of phosphorylated ERK1/2 and Akt protein expressions showed an increase, as compared to the corresponding amount in the control group. On the other hand, the amount of phosphorylated p38MAPK protein expression showed a decrease from the amount in the control group after the addition of MIP-1alpha. U0126 (a MEK1/2 inhibitor) or LY294002 (a PI3K inhibitor) was added to ST2 and MC3T3-E1 cells, and was found to inhibit RANKL mRNA and RANKL protein expression in these cells. When SB203580, a p38MAPK inhibitor, was added, RANKL mRNA and RANKL protein expression were increased in these cells. MIP-1alpha was found to promote osteoclastic differentiation of C7 cells, an osteoclastic precursor cell line, in a MIP-1alpha concentration-dependent manner. MIP-1alpha promoted differentiation into osteoclasts more extensively in C7 cells incubated together with ST2 and MC3T3-E1 cells than in C7 cells incubated alone. These results suggested that MIP-1alpha directly acts on the osteoclastic precursor cells and induces osteoclastic differentiation. This substance also indirectly induces osteoclastic differentiation through the promotion of RANKL expression in bone-marrow stromal cells and osteoblasts. The findings of this investigation suggested that activation of the MEK/ERK and the PI3K/Akt pathways and inhibition of p38MAPK pathway were involved in RANKL expression induced by MIP-1alpha in bone-marrow stromal cells and osteoblasts. This finding may be useful in the development of an osteoclastic inhibitor that targets intracellular signaling factors.
  • Immature and mature neurons coexist among glial scars after rat traumatic brain injury., Tatsuki Itoh, Takao Satou, Shozo Nishida, Shigeo Hashimoto, Hiroyuki Ito, Neurological research, Neurological research, 29(7), 734 - 42, Oct. 2007 , Refereed
    Summary:OBJECTIVES: Glial scars around a damaged area after brain injury inhibit neurite elongation from surviving neurons and axonal plasticity, and thus prevent neural network regeneration. However, the generation, differentiation and maturation of neural stem cells (NSCs) among glial scars after brain injury have not yet been reported. METHODS: In the present study, we investigated the chronological relationship between gliosis and maturation of new neurons around a damaged area using a rat traumatic brain injury (TBI) model. RESULTS: Between 1 and 7 days after injury, many nestin-positive cells were observed around the damaged area. Three days after injury, many small nestin-positive cells showed an astrocytic morphology. Between 1 and 30 days after injury, doublecortin (DCX)-positive cells were present around the damaged area. Three and 7 days after injury, a small number of nestin-positive cells were immunopositive for glial fibrillary acidic protein (GFAP). Seven days after injury, there were DCX-positive cells in the gliosis occurring in the lesion. Thirty days after injury, DCX-positive cells were observed near and among the glial scars and a small number of these cells were immunopositive for NeuN. DISCUSSION: These results suggest that DCX-positive cells were present near and among the glial scars after brain injury, and that these cells changed from immature to mature neurons. It is considered that promotion of the maturation and differentiation of newly formed immature neurons near and among glial scars after injury may improve the brain dysfunction induced by glial scars after brain injury.
  • The protein kinase C inhibitor, H7, inhibits tumor cell invasion and metastasis in mouse melanoma via suppression of ERK1/2., Masanobu Tsubaki, Hiroshi Matsuoka, Chikako Yamamoto, Chisato Kato, Mitsuhiko Ogaki, Takao Satou, Tatsuki Itoh, Takashi Kusunoki, Yoshihiro Tanimori, Shozo Nishida, Clinical & experimental metastasis, Clinical & experimental metastasis, 24(6), 431 - 8, 2007 , Refereed
    Summary:Protein kinase C (PKC) has been shown to be a signal transducer during tumorigenesis, tumor cell invasion, and metastasis. Recent studies have reported that the PKC inhibitor, 7-hydroxystaurosporine, inhibits tumor cell invasion. However, the molecular mechanisms of this inhibition of invasion and metastasis are not well understood. In the present study, we attempt to clarify the mechanism by which H7, a PKC inhibitor, inhibits tumor cell invasion and metastasis in the melanoma cell line B16BL6. It was found that H7 inhibits B16BL6 cell invasion and metastasis. We also observed that H7 inhibits the mRNA expression and protein activities of matrix metalloproteinase (MMP)-1, -2, -9 and MT1-MMP. Furthermore, H7 suppresses phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2). However, other signal transduction factors, such as p38 mitogen-activated protein kinase (p38MAPK) and c-Jun N-terminal kinase 1/2 (JNK1/2), were unaffected. Moreover, U0126, a MEK1/2 inhibitor, also inhibited B16BL6 cell invasion and metastasis, as well as the mRNA expression and protein activities of MMP-1, -2, -9 and MT1-MMP. This indicates that H7 inhibits signal transduction through the PKC/MEK/ERK pathway, thereby inhibiting B16BL6 cell invasion and metastasis. These results suggest that PKC inhibitors have potential clinical applications in the treatment of tumor cell metastasis.
  • Cultured rat astrocytes give rise to neural stem cells., Tatsuki Itoh, Takao Satou, Shozo Nishida, Shigeo Hashimoto, Hiroyuki Ito, Neurochemical research, Neurochemical research, 31(11), 1381 - 7, Nov. 2006 , Refereed
    Summary:Previously, we reported the occurrence of neural stem cells (NSCs) around an area of damage after rat traumatic brain injury (TBI), but it was unclear if this was due to blastgenesis in astrocytes, or to NSCs migrating from the subventricular zone (SVZ). In this study, NSCs were isolated and cultured from cultured type 1 astrocytes taken from newborn rat cortex in which the subventricular zone and hippocampus had been discarded. All cultured type 1 astrocytes showed glial fibrillary acidic protein (GFAP) immunopositivity. Nestin immunopositive spheres were isolated from type 1 astrocytes and cultured in the presence of bFGF and EGF in the medium. Neurospheres differentiated into Tuj1-, GFAP- and A2B5-positive cells after 4 days of culture without bFGF and EGF. These results indicate that isolated neurospheres from brain cortex astrocytes can differentiate into neurons and glia and might contribute to neurogenesis and neuroplasticity.
  • Nitrogen-containing bisphosphonate, YM529/ONO-5920 (a novel minodronic acid), inhibits RANKL expression in a cultured bone marrow stromal cell line ST2., Shozo Nishida, Masanobu Tsubaki, Mayumi Hoshino, Ayumi Namimatsu, Hiromi Uji, Shohei Yoshioka, Yoshihiro Tanimori, Masashi Yanae, Masahiro Iwaki, Kiyohiro Irimajiri, Biochemical and biophysical research communications, Biochemical and biophysical research communications, 328(1), 91 - 7, Mar. 04 2005 , Refereed
    Summary:Increase in bone resorption by osteoclasts can cause metabolic bone diseases, such as osteoporosis. Recent attention has been paid to the receptor activator of the NF-kappaB ligand (RANKL), an accelerator of osteoclast differentiation. RANKL is expressed on the bone marrow-derived stromal cell membrane and induces the differentiation of osteoclasts by binding to RANK expressed on the osteoclast precursor cell membrane. Since the inhibition of RANKL expression can lead to the inhibition of osteoclastic bone resorption, the clinical application of RANKL inhibition could be expected to have a major effect on metabolic bone disease therapy. In this study, we investigated whether or not YM529/ONO-5920, a nitrogen-containing bisphosphonate (a novel minodronic acid), inhibits RANKL expression in a bone marrow-derived stromal cell line (ST2 cells). Reverse transcription-polymerase chain reaction revealed that the administration of YM529/ONO-5920 to ST2 cells inhibited RANKL mRNA expression and reduced RANKL proteins as assessed by Western blot analysis. The inhibition of RANKL mRNA expression was reversed when geranylgeranyl pyrophosphate (GGPP), an intermediate in the mevalonate pathway, was used in combination. Furthermore, YM529/ONO-5920 reduced phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2), and similarly, U0126, a mitogen-activated protein kinase kinase 1/2 inhibitor, inhibited RANKL expression. Pretreatment with GGPP reversed the YM529/ONO-5920-induced decrease in phosphorylation of ERK. Furthermore, YM529/ONO-5920 decreased TRAP-positive cells in co-culture of ST2 cells and an osteoclast cell line, C7 cells, and this decrease was inhibited by pretreatment with GGPP. This indicates that YM529/ONO-5920 inhibits GGPP biosynthesis in the mevalonate pathway and then signal transduction in the Ras-mitogen-activated protein kinase pathway, thereby inhibiting RANKL expression on ST2 cells. These results suggest a newly elucidated action of bisphosphonates in the inhibition of bone resorption.
  • Scanning electron microscope study of the rabbit anterior chamber angle., Shozo Nishida, Hideya Uchida, Makoto Takeuchi, Guii Qin Sui, Satoshi Mizutani, Masayoshi Iwaki, Medical molecular morphology, Medical molecular morphology, 38(1), 54 - 62, Mar. 2005 , Refereed
    Summary:The three-dimensional fine structure and the collagenous framework of the anterior chamber angle (chamber angle) of albino rabbits, which does not posses a Schlemm's canal, were examined, with or without NaOH maceration, by scanning electron microscopy. The chamber angle is ill defined because the deep ciliary cleft appears parceled by iris pillars. The collagenous framework of the pectinate ligaments and iris pillars arising from the iris root join to the collagen bundles of the trabecular sheet, and the surface of the ciliary cleft is covered by an endothelial cell layer that screens the trabecular meshwork. Round openings are apparent in the recesses of the ciliary cleft, with junctions between more superficial or deeper trabecular sheets resulting in direct communication between adjacent trabecular spaces. Openings of the angular aqueous plexus/sinus in place of Schlemm's canal were seen on the inner surface of the corneosclera after removing the trabecular meshwork. The framework of the trabecular sheet is basically formed of circumferentially arranged flat collagen bundles and reticularly diverged narrow collagen bundles extending from them. There is no shift between collagen bundles of the trabecular meshwork and the corneosclera, suggesting a frail connection between the two tissues.
  • Anti-proliferative effect of Ginseng Radix on human premyelocytic leukemia cells (HL-60), Hideaki Matsuda, Noriko Hirata, Shunsuke Naruto, Shozo Nishida, Kiyohiro Irimajiri, Keiichi Samukawa, Michinori Kubo, Journal of Traditional Medicines, Journal of Traditional Medicines, 22(1), 4 - 8, 2005 , Refereed
    Summary:Anti-proliferative effect of Ginseng Radix on human premyelocytic leukemia cells (HL-60) was investigated. The 70% methanol extract (RG-ext) of Red Ginseng (steamed and dried roots of Panax ginseng C. A. MEYER) had superior activity to that (WG-ext) of White Ginseng (peeled and dried root of P. ginseng). Among major saponins of Ginseng Radix, ginsenoside-Rc (G-Rc) and -Rb2(G-Rb2) exhibited significant activity, but ginsenoside-Rb1 (G-Rb1), -Rg1 (G-Rg1), -Rg2 (G-Rg2) and -Ro (G-Ro) were ineffective. (20S)-Ginsenoside-Rg3[(20S)-G-Rg3] formed by the processing of Red Ginseng from the crude root of P. ginseng showed remarkable activity. Based on the morphological changes and the pattern of DNA fragmentation, RG-ext, (20S)-G-Rg3, G-Rb2, and G-Rc were found to induce apoptosis in HL-60 cells. Additionally, the effect of (20S)-G-Rg3 was enhanced by treatment together with a water-soluble non-saponin fraction (RG-ext-W) obtained from RG-ext. © 2005, Medical and Pharmaceutical Society for WAKAN-YAKU. All rights reserved.
  • Mevastatin induces apoptosis in HL60 cells dependently on decrease in phosphorylated ERK., Shozo Nishida, Hiroshi Matsuoka, Masanobu Tsubaki, Yoshihiro Tanimori, Masasi Yanae, Yoshiki Fujii, Masahiro Iwaki, Molecular and cellular biochemistry, Molecular and cellular biochemistry, 269(1-2), 109 - 14, Jan. 2005 , Refereed
    Summary:Mevastatin which is an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme in cholesterol synthesis, suppress cell proliferation and induce apoptosis. However, the molecular mechanism of apoptosis induction is not well understood. So, in the present study, we attempted to clarify the mechanism by which mevastatin induces apoptosis in HL60 cells. It was found that mevastatin induced apoptosis. At that time, we observed an increase in caspase-3 activity and morphological fragmentation of the nuclei. The apoptosis induced by mevastatin was not inhibited by the addition of farnesyl pyrophosphate (FPP), squalene, ubiquinone, and isopentenyladenine, but was inhibited by the addition of geranylgeranyl pyrophosphate (GGPP). When we examined the survival signals at the time of apoptotic induction, we also observed that the administration of mevastatin had caused a remarkable decrease in the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). However, other survival signals, such as nuclear factor kappa B (NF-kappaB), protein kinase B (Akt), and p38 mitogen-activated protein kinase (p38), exhibited no change. In addition, no quantitative change was observed in Bcl-2, which was an anti-apoptosis protein. It was also observed that apoptosis was induced when U0126, an MEK inhibitor, was added to the cells to inhibit ERK. These results suggested that mevastatin induced apoptosis when it inhibited GGPP biosynthesis and consequently decreased the level of phosphorylated ERK, which was a survival signal; moreover, at that time, there was no influence on NF-kappaB, Akt, p38, and Bcl-2. The results of this study also suggested that mevastatin could be used as an anticancer agent.
  • Diethyldithiocarbamate can induce two different type of death: apoptosis and necrosis mediating the differential MAP kinase activation and redox regulation in HL60 cells., Saori Kimoto-Kinoshita, Shozo Nishida, Takanori T Tomura, Molecular and cellular biochemistry, Molecular and cellular biochemistry, 265(1-2), 123 - 32, Oct. 2004 , Refereed
    Summary:Although apoptosis and necrosis have been considered different pathways to cell death, only one compound induces both types of cell death. Diethyldithiocarbamate (DDC) has been shown to have antioxidant or prooxidant effects in several different systems. We observed in our present study that DDC induced not only apoptosis but also necrosis depending on its dosage in HL60 premyelocytic leukemia cells. Moreover, in hypoxia cell culture conditions, DDC-induced necrotic cells decreased but DDC-induced apoptosis continued. We investigated the DDC-induced different cell death mechanisms as they are correlated with reactive oxygen species (ROS). High-dose DDC-induced necrotic cell death is thought to depend on the increase of intracellular ROS, while low-dose DDC-induced apoptosis is thought to depend on changes of the intracellular redox state by the transporting of external metal ions. There was no sequential or quantitative change of Bcl-2 family proteins in DDC-induced apoptotic or necrotic pathways. However, the mitochondrial transmembrane potential was remarkably decreased in the DDC-induced necrosis. Finally, duration of c-Jun N-terminal kinase (JNK) activation resulted in different types of cell death.
  • Pretreatment with PKC inhibitor triggers TNF-alpha induced apoptosis in TNF-alpha-resistant B16 melanoma BL6 cells., Shozo Nishida, Shohei Yoshioka, Saori Kinoshita-Kimoto, Michiyo Kotani, Masanobu Tsubaki, Yoshiki Fujii, Takanori T Tomura, Kiyohiro Irimajiri, Life sciences, Life sciences, 74(6), 781 - 92, Dec. 26 2003 , Refereed
    Summary:Tumor necrosis factor alpha (TNF-alpha) modulates various events through several different pathways. Many tumor cells are resistant to this cytokine. Pretreatment of these cells with actinomycin D enhances TNF-alpha-induced apoptosis. In the present study, we investigated the mechanism of this enhancement and whether or not the apoptosis of TNF-alpha-resistant cancer cells can be induced by the inhibition of Protein kinase C (PKC). When TNF-alpha was added after inhibition of PKC by H7, apoptosis was observed, and companied with the activation of nuclear factor kappa B (NF-kappaB). After the inhibition of protein kinase B (Akt) by LY294002 or p38 mitogen-activated protein kinase (p38MAPK) by SB203580, the addition of TNF-alpha did not cause apoptosis. However, after the inhibition of MAPK/extracellular signal-regulated kinase kinase 1/2 (MEK1/2) with U0126, apoptosis was observed when TNF-alpha was added. In the Western blotting analysis, phosphorylation of MEK1/2 occurred at 60 minutes after the addition of TNF-alpha. However, it was noted that after pretreatment with H7, a significant decrease in phosphorylated MEK1/2 was observed. The present findings suggest that MEK1/2 plays an important role in TNF-alpha-resistance in TNF-alpha-resistant B16 melanoma BL6 cells. Furthermore, it was found that MEK1/2 is more important than NF-kappaB, Akt, and p38MAPK in anti-apoptotic PKC signaling and that TNF-alpha-resistance can be overcome by inhibiting MEK1/2. These results suggest the possibility of development of a new anticancer drug treatment.
  • A new bisphosphonate, YM529 induces apoptosis in HL60 cells by decreasing phosphorylation of single survival signal ERK., Shozo Nishida, Yoshiki Fujii, Shohei Yoshioka, Sigeru Kikuichi, Masanobu Tsubaki, Kiyohiro Irimajiri, Life sciences, Life sciences, 73(21), 2655 - 64, Oct. 10 2003 , Refereed
    Summary:It is believed that bisphosphonates (BPs) induce apoptosis in cells such as myeloma cells, as they inhibit prenylation of G-proteins. However, the details of the apoptosis-inducing mechanism remain obscure. In the present study, we attempted to clarify the mechanism by which YM529, a new bisphosphonate, induces apoptosis. YM529 induced cell deaths in HL60 cells in a concentration-dependent manner. At that time, we observed an increase in Caspase-3 activity and morphological fragmentation of the nuclei. We could confirm that these cell deaths were evidence of apoptosis. The apoptosis induced by YM529 was not inhibited by the addition of farnesyl pyrophosphate (FPP), but was by the addition of geranylgeranyl pyrophosphate (GGPP). When we examined the survival signals at the time of apoptotic induction, we also observed that the administration of YM529 caused a remarkable decrease in the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). However, other survival signals such as nuclear factor kappa B (NF-kappaB), protein kinase B (Akt), and p38 mitogen-activated protein kinase (p38) exhibited no change. In addition, no quantitative change was observed in Bcl-2, which is an anti-apoptosis protein. It was also observed that apoptosis was induced when U0126, an MEK inhibitor, was added to the cells to inhibit ERK. These results suggest that YM529, the new bisphosphonate, induced apoptosis when inhibit GGPP synthase and consequently decreased the levels of phosphorylated ERK, which is a survival signal; moreover, during this process, there is no influence on NF-kappaB, Akt, p38, and Bcl-2. The results of this study also suggest that YM529 can be used as an anticancer agent, in addition to its use as a therapeutic agent to treat osteoporosis.
  • Induction of apoptosis in HL-60 cells treated with medicinal herbs, S Nishida, S Kikuichi, S Yoshioka, M Tsubaki, Y Fujii, H Matsuda, M Kubo, K Irimajiri, AMERICAN JOURNAL OF CHINESE MEDICINE, AMERICAN JOURNAL OF CHINESE MEDICINE, 31(4), 551 - 562, 2003 , Refereed
    Summary:In order to develop a new apoptosis inducer, we screened 22 crude drugs for their apoptosis-inducing activity. It was found that Glycyrrhiza uralensis, Cynomorium songaricum, Eucommia ulmoides, Phellodendron amurense, Cinnamomum cassia and Paeonia lactiflora induced the death of HL-60 cells. To investigate the mechanism of apoptosis induced by these six crude drugs, the mitochondrial transmembrane potential and the activity of caspase-3 were measured. Reduced mitochondrial transmembrane potentials within 12 hours after the administration of Glycyrrhiza uralensis, Cynomorium songaricum, Phellodendron amurense and Paeonia lactiflora, and within 24 hours after the administration of Eucommia ulmoides and Cinnamomum cassia were observed. All of the six apoptosis-inducing crude drugs increased caspase-3 activity within 12-36 hours after administration. After further examining the apoptosis-inducing activity of berberine, palmatine, panelofuroline and glycyrrhizin, which were the ingredients obtained from Phellodendron amurense, Glycyrrhiza uralensis and Paeonia lactiflora, it was found that only berberine could induce apoptosis. From these results, it was concluded that the apoptosis induced by the six crude drugs (Glycyrrhiza uralensis, Cynomorium songaricum, Eucommia ulmoides, Phellodendron amurense, Cinnamomum cassia and Paeonia lactiflora) occurred via the mitochondrial route and that the apoptosis-conducting mechanism acted through a cascade involving caspase-3.
  • Apoptosis-inducing effect of a new bisphosphonate, YM529, on various hematopoietic tumor cell lines., Shozo Nishida, Shigeru Kikuichi, Hisae Haga, Shohei Yoshioka, Masonobu Tsubaki, Katsuki Fujii, Kiyohiro Irimajiri, Biological & pharmaceutical bulletin, Biological & pharmaceutical bulletin, 26(1), 96 - 100, Jan. 2003 , Refereed
    Summary:In recent years, it has been reported that bisphosphonates inhibited the cell cycle of myeloma cells to inhibit cell proliferation directly, and it was also reported that bisphosphonates induced apoptosis of myeloma cells in vitro. Recently, YM529 was developed as a new third-generation bisphosphonate. In our experiment, we investigated whether YM529 showed an antitumor effect on hematopoietic tumor cell lines other than myeloma, and we compared YM529 with YM175, which had a relatively more potent antitumor effect than that of existing bisphosphonates. We found that YM529 inhibited cell proliferation in various hematopoietic tumor cell lines (acute promyelocytic leukemia cell line HL-60, chronic myeloid leukemia cell line K562, histiocytic lymphoma cell line U937, lymphoblastic leukemia T cell line Jurkat, acute lymphoblastic leukemia T cell line MOLT-4, lymphoblastic leukemia B cell line CCRF-SB) including myeloma (myeloma cell line HS-Sultan) dose-dependently and time-dependently to a degree equivalent or superior to that in myeloma, and induced apoptosis at a lower concentration as compared with YM175. We confirmed many dead cells as well as apoptosis based on the detection of the nuclei with separate globular structure, the activation of caspase-3, and the decrease in mitochondrial transmembrane potential. Therefore, it is concluded that further utilization of YM529 can be expected against hematopoietic tumor cells in the future.
  • Type IV collagen, type IV collagenase activity and ability of cell proliferation in human thyroid tumours., Takeshi Kusunoki, Shozo Nishida, Saori Kimoto-Kinoshita, Kiyotaka Murata, Takao Satou, Takanori Tomura, Asian journal of surgery, Asian journal of surgery, 25(4), 304 - 8, Oct. 2002 , Refereed
    Summary:BACKGROUND: The processes of malignant tumour invasion and metastasis are known to include the destruction of cell stroma and vascular basement membrane. It has been suggested that type IV collagenase degrades type IV collagen, a main component of the basement membrane. METHODS: In our study, type IV collagenase activity in human thyroid tumours was measured by the Liotta method. The degree of destruction of diseased regions of thyroid tumours was immunohistochemically determined by anti-type IV collagen antibody staining. Cell proliferation in the tumours was estimated using anti-proliferating cell nuclear antigen (PCNA) and epidermal growth factor receptor (EGFR). RESULTS: T4 thyroid carcinomas with higher type IV collagenase activity and very weak type IV discontinuous immunostaining for type IV collagen of follicular basement membranes, exhibited many PCNA or EGFR positive cells. In benign tumours, normofollicular- or macrofollicular-type tumours with low type IV collagenase activity showed few PCNA and EGFR positive cells and intact type IV collagen of basement membranes, as seen in normal thyroids. Conversely, an atypical adenoma with higher type IV collagenase activity showed many PCNA and EGFR positive cells and weak type IV discontinuous immunostaining for type IV collagen, as in thyroid carcinomas. CONCLUSION: These findings suggest that staining for type IV collagen and type IV collagenase activity reflect the ability of cell proliferation, and help predict the aggressiveness of invasion and metastasis in human thyroid tumours.
  • Histopathological findings of human thyroid tumors and dynamic MRI., Takeshi Kusunoki, Kiyotaka Murata, Shozo Nishida, Takanori Tomura, Masaaki Inoue, Auris, nasus, larynx, Auris, nasus, larynx, 29(4), 357 - 60, Oct. 2002 , Refereed
    Summary:OBJECTIVE: Time intensity curves for gadolinium-diethylenetriaminepentaceticacid (Gd-DTPA) enhanced magnetic resonance imaging (MRI), namely dynamic MRI, were determined for thyroid diseases, and compared with findings of histopathologic examination. METHODS: Time intensity curves for the thyroid tumors were determined. Three different patterns of time intensity curves were observed: a rapid washout pattern, a delayed washout pattern and no change. Cell proliferating activities of thyroid tumors were estimated immunohistochemically by proliferating cell nuclear antigen (PCNA). RESULTS: In most of malignant diseases and a few benign diseases that had marked cell proliferative activity with showing more than 20% on labeling indexes of PCNA, the time intensity curve displayed the delayed washout pattern, in which intensity was above half-maximal value within 10 min after injection Gd-DTPA. Almost all benign diseases and a few well differentiated carcinomas displayed the rapid washout pattern, in which intensity was decreased to lower than half of peak grade within 10 min following injection and showed from 10 to 20% on labeling indexes of PCNA. Benign diseases that displayed no change of time intensity curve did not show PCNA positive cells. CONCLUSION: These findings suggested that the time intensity curve obtained from dynamic MRI might indicate cell proliferating activity of thyroid tumors.
  • Change of Cu,Zn-superoxide dismutase activity of guinea pig lung in experimental asthma., Shozo Nishida, Kazuhiro Teramoto, Saori Kimoto-Kinoshita, Yuji Tohda, Shigenori Nakajima, Takanori T Tomura, Kiyohiro Irimajiri, Free radical research, Free radical research, 36(6), 601 - 6, Jun. 2002 , Refereed
    Summary:Correlation between the level of reactive oxygen species (ROS) generated by airway inflammatory cells and superoxide dismutase (SOD) activity of pulmonary tissue during an asthma attach was investigated in a guinea pig model of allergic asthma. In addition, the influence of SOD inhibition by diethyldithiocarbamate (DDC, Cu-chelating agent) on the airway was investigated in terms of pulmonary function during an asthma attach. Relative to controls, the capacity of bronchoalveolar lavage fluid (BAL) cells to release ROS was significantly increased in guinea pigs sensitized with ovalbumin (OA) as the antigen, and significantly increased in guinea pigs with an asthma attack provoked by the inhalation of OA. SOD activity was increased significantly in the antigen-sensitized group. The asthma provocation group showed a tendency for increase in total SOD activity, compared with the sensitization group, whose increase was dependent on the increase in copper, zinc-SOD (Cu, Zn-SOD) activity. Pretreatment with DDC increased the severity and duration of the asthma attack. These results were indicated that Cu, Zn-SOD was closely involved in the asthma process, particularly in the scavenging of oxygen radicals secreted from BAL cells.
  • Correlation between morphology of colonies formed by human thyroid tumor cells in collagen gel culture and invasive ability., Takeshi Kusunoki, Shozo Nishida, Kiyotaka Murata, Takanori Tomura, Thyroid : official journal of the American Thyroid Association, Thyroid : official journal of the American Thyroid Association, 12(4), 281 - 6, Apr. 2002 , Refereed
    Summary:Two different types of colonies from human thyroid papillary carcinomas were observed in collagen gel culture. One was a branching type, with many three-dimensional outgrowths. The other was a spherical type, with no outgrowths. As viewed by electron microscopy, the tumor cells of spherical-type colonies had more microvilli on their surfaces than did cells in the branching-type colonies. Histochemically, the tumor cells of spherical-type colonies had stronger positive reactions than did tumor cells of branching-type colonies. This suggests that tumor cells of spherical-type colonies have a higher potential for dissemination and invasion than do tumor cells of branching-type colonies. In fact, cases with higher percentages of spherical-type colonies could show extracapsular invasion histopathologically. Our results indicate that collagen gel culture is a useful method for studying the heterogeneity of tumor cells as well as their invasive ability in vitro.
  • Protease-activated receptor-2 (PAR-2) in the rat gastric mucosa: immunolocalization and facilitation of pepsin/pepsinogen secretion., Naoyuki Kawao, Yuriko Sakaguchi, Ai Tagome, Ryotaro Kuroda, Shozo Nishida, Kiyohiro Irimajiri, Hiroyuki Nishikawa, Kenzo Kawai, Morley D Hollenberg, Atsufumi Kawabata, British journal of pharmacology, British journal of pharmacology, 135(5), 1292 - 6, Mar. 2002 , Refereed
    Summary:1. Agonists of protease-activated receptor-2 (PAR-2) trigger neurally mediated mucus secretion accompanied by mucosal cytoprotection in the stomach. The present study immunolocalized PAR-2 in the rat gastric mucosa and examined if PAR-2 could modulate pepsin/pepsinogen secretion in rats. 2. PAR-2-like immunoreactivity was abundant in the deep regions of gastric mucosa, especially in chief cells. 3. The PAR-2 agonist SLIGRL-NH(2), but not the control peptide LSIGRL-NH(2), administered i.v. repeatedly at 0.3 - 1 micromol kg(-1), four times in total, significantly facilitated gastric pepsin secretion, although a single dose produced no significant effect. 4. The PAR-2-mediated gastric pepsin secretion was resistant to omeprazole, N(G)-nitro-L-arginine methyl ester (L-NAME) or atropine, and also to ablation of sensory neurons by capsaicin. 5. Our study thus provides novel evidence that PAR-2 is localized in mucosal chief cells and facilitates gastric pepsin secretion in the rats, most probably by a direct mechanism.
  • Morphological and functional differentiation of human thyroid cells in collagen gel culture, T Kusunoki, S Nishida, M Koezuka, K Murata, T Tomura, AURIS NASUS LARYNX, AURIS NASUS LARYNX, 28(4), 333 - 338, Nov. 2001 , Refereed
    Summary:Objective: cultured human thyroid cells in collagen gel culture were examined on cell morphology and the production of thyroglobulin (Tg), triiodothyronine (T3) and thyroxine (T4) which are components of functional differentiation. Methods: thyroid cells obtained from normal human thyroid tissues (four cases), follicular adenoma tissues (three cases). papillary carcinoma tissues (three cases), and follicular carcinoma tissue (one case), were cultured in collagen gel. Then these cultured cells were observed on cellular morphology and production of Tg, T3 and T4. Moreover, changes in morphological characteristics and production of Tg, T3 and T4 induced by addition of thyroid stimulating hormone (TSH) and epidermal growth factor (EGF) to medium in collagen gel culture were determined. Results: normal and tumor cells in collagen gel culture formed colonies and follicles with Tg production, similar to in vivo-like three-dimensional cellular structures and functions. Normal thyroid cells stimulated TSH induced more Tg and produced morphological changes, i.e. enlarged follicular lumens and increased the height of follicular cells, but did not promote cell proliferation. Reversely, normal thyroid cells stimulated with EGF promoted cell proliferation, but did not change morphological findings and did not increase production of Tg, T3 and T4, Conclusion: these findings suggest that collagen gel culture is useful for observing the effects of stimulation by cell growth factor on the morphological and functional differentiation of human thyroid cells. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.
  • Cathepsin L activity and its inhibitor in human otitis media, T Kusunoki, S Nishida, K Murata, K Kobashi, H Nakatani, T Hiwasa, T Tomura, JOURNAL OF OTOLARYNGOLOGY, JOURNAL OF OTOLARYNGOLOGY, 30(3), 157 - 161, Jun. 2001 , Refereed
    Summary:Objective: We examined cathepsin L activity, expression of cystatin A, and copper- and zinc-containing superoxide dismutase in human chronic otitis media. The relationships of our findings to clinical findings (e.g., grade of bone destruction) were also studied. Design: Retrospective basic and clinical study. Setting: Department of Otolaryngology and First Department of Biochemistry, Kinki University School of Medicine, Osaka, Japan. Method: The human middle ear tissues evaluated in this study were surgically obtained from seven patients with cholesteatoma epithelium, three patients with granulation tissues in cholesteatoma, three patients with granulation tissues in noncholesteatoma, and three patients with intact mucous membrane of the middle ear. Main Outcome Measures: Cathepsin L activities in cholesteatoma epithelium, granulation tissues in cholesteatoma, or granulation tissues in noncholesteatoma were measured using Barrett's method. Cystatin A expressions were observed by Western blot analysis. Copper- and zinc-containing superoxide dismutase in cholesteatoma was examined immunohistochemically. Results: Mean cathepsin L activity was higher in diseased tissues than in intact mucous membranes of the middle ear. Granulation tissues with high cathepsin L activity resulted in extensive bone destruction in both cholesteatomas and non-cholesteatomas of the middle ear. All cases with intact mucous membrane of the middle ear exhibited no expression of cystatin A. Seven of 10 cases with diseased tissues expressed cystatin A in cholesteatoma epithelium, granulation tissues in cholesteatoma, or granulation tissues in noncholesteatoma. No relationships were found between cystatin A expression and grade of cathepsin L activity. Copper- and zinc-containing superoxide dismutase was more strongly positive in cholesteatoma epithelium regions than in granulation tissues. Conclusion: These results suggest that copper- and zinc-containing superoxide dismutase in cholesteatoma epithelium prevents complications by suppressing cathepsin L activity.
  • Decrease of endothelial nitric oxide synthase in stroke-prone spontaneously hypertensive rat cerebral cortex, S Kimoto-Kinoshita, S Nishida, TT Tomura, NEUROSCIENCE LETTERS, NEUROSCIENCE LETTERS, 288(2), 103 - 106, Jul. 2000 , Refereed
    Summary:We reported in our previous study that constitutive nitric oxide synthase (cNOS) activity significantly decreased and oxygen stress increased in the 31-week-old stroke-prone spontaneously hypertensive rat (SHRSP) cerebral cortex (CC). In the present study we examined the protein amount of two cNOS isoforms, the neuronal and the endothelial types, in SHRSP CC using Western blot analysis. Although no significant difference was observed in the amount of neuronal NOS (nNOS) protein, endothelial NOS (eNOS) protein prominently decreased in 31-week-old SHRSP CC compared to age-matched Wistar Kyoto rat and 15-week-old SHRSP. In rats at this age, we also observed a large quantity of albumin in the protein amount. However, the protein amount of heat shock protein 70, which is a molecular chaperon and a marker of injury, showed no significant changes. These results indicate that the alteration of eNOS but not of nNOS protein would be more closely associated with the development of stroke in SHRSP. (C) 2000 Published by Elsevier Science Ireland Ltd.
  • Type IV collagenase and immunostaining of type IV collagen in human thyroid tumors, Takeshi Kusunoki, Shozo Nishida, Saori Kimoto-Kinoshita, Kiyotaka Murata, Takao Satou, Takanori Tomura, Auris Nasus Larynx, Auris Nasus Larynx, 27(2), 161 - 165, Apr. 2000
    Summary:Objective: Required stages in the processes of malignant tumor invasion and metastasis are known to include the destruction of cell stroma and vascular basement membrane. It has been suggested that type IV collagenase can degrade type IV collagen, a main component of basement membrane. In our study, type IV collagenase activity was compared with the grade of destruction of type IV collagen by thyroid tumors. Methods: Type IV collagenase activity was measured in human thyroid tissue obtained surgically from four patients with Graves' disease, four with follicular adenoma, six with papillary carcinoma, and four with follicular carcinoma. Six normal thyroid tissue specimens were also studied. The grade of destruction in the diseased regions of thyroid tumors and surrounding intact tissues were determined immunochemically by anti-type IV collagen antibody staining. Results: Tumors with high type IV collagenase activity exhibited extracapsular invasion, lymph node metastasis and very weak discontinuity of immunostaining for type IV collagen from the basement membrane in diseased regions. In addition, surrounding intact tissues exhibited weak immunostaining for type IV collagen. Tumors with low type IV collagenase activity exhibited neither extracapsular invasion nor lymph node metastasis, but did exhibit weak immunostaining for type IV collagen from the basement membranes in diseased regions. However, the surrounding intact tissues had preserved type IV collagen in follicular basement membranes. Conclusion: These findings suggest that staining for type IV collagen can be considered a biochemical marker for prediction of the aggressiveness of invasion and metastasis. (C) 2000 Elsevier Science Ireland Ltd.
  • Type IV collagenase and immunostaining of type IV collagen in human thyroid tumors, Takeshi Kusunoki, Shozo Nishida, Saori Kimoto-Kinoshita, Kiyotaka Murata, Takao Satou, Takanori Tomura, Auris Nasus Larynx, Auris Nasus Larynx, 27(2), 161 - 165, Apr. 2000 , Refereed
    Summary:Objective: Required stages in the processes of malignant tumor invasion and metastasis are known to include the destruction of cell stroma and vascular basement membrane. It has been suggested that type IV collagenase can degrade type IV collagen, a main component of basement membrane. In our study, type IV collagenase activity was compared with the grade of destruction of type IV collagen by thyroid tumors. Methods: Type IV collagenase activity was measured in human thyroid tissue obtained surgically from four patients with Graves' disease, four with follicular adenoma, six with papillary carcinoma, and four with follicular carcinoma. Six normal thyroid tissue specimens were also studied. The grade of destruction in the diseased regions of thyroid tumors and surrounding intact tissues were determined immunochemically by anti-type IV collagen antibody staining. Results: Tumors with high type IV collagenase activity exhibited extracapsular invasion, lymph node metastasis and very weak discontinuity of immunostaining for type IV collagen from the basement membrane in diseased regions. In addition, surrounding intact tissues exhibited weak immunostaining for type IV collagen. Tumors with low type IV collagenase activity exhibited neither extracapsular invasion nor lymph node metastasis, but did exhibit weak immunostaining for type IV collagen from the basement membranes in diseased regions. However, the surrounding intact tissues had preserved type IV collagen in follicular basement membranes. Conclusion: These findings suggest that staining for type IV collagen can be considered a biochemical marker for prediction of the aggressiveness of invasion and metastasis. (C) 2000 Elsevier Science Ireland Ltd.
  • Age-related change of antioxidant capacities in the cerebral cortex and hippocampus of stroke-prone spontaneously hypertensive rats, S Kimoto-Kinoshita, S Nishida, TT Tomura, NEUROSCIENCE LETTERS, NEUROSCIENCE LETTERS, 273(1), 41 - 44, Sep. 1999 , Refereed
    Summary:Regional distribution and age-related change of Mn-, Cu/Zn-superoxide dismutase (SOD) and constitutive type of nitric oxide synthase (NOS) activities in the brain were determined using stroke-prone spontaneously hypertensive rats (SHRSP). In the hippocampus (HIP), Mn- and Cu/Zn-SOD activities in SHRSP of 31-week-old were significantly lower than those of 15-week-old or normotensive rats (WKY). From Mn-SOD immunohistochemical staining of several subfields of the HIP, our results suggested that SHRSP hippocampal CA1 was more vulnerable to oxidative stress compared with WKY and other subfields. In the 31-week-old SHRSP cerebral cortex (CC), the activities of Mn-, Cu/Zn-SOD and NOS were significantly lower than those in WKY. At this age, most of the SHRSP developed cerebral injuries. These observations indicated that hypertensive vascular disease observed in the SHRSP CC resulted from the decreased antioxidant capacity that is closely associated with the development of stroke and, in turn, shortened life span. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.
  • Malignancies of human thyroid tumors and dynamic magnetic resonance imaging (MRI), T. Kusunoki, K. Murata, H. Hosoi, S. Nishida, T. Tomura, M. Inoue, Auris Nasus Larynx, Auris Nasus Larynx, 25(4), 419 - 424, Dec. 1998 , Refereed
    Summary:Time intensity curves for gadolinium-diethylene triaminepentacetic acid (Gd-DTPA) enhanced magnetic resonance imaging (MRI), namely dynamic MRI, were determined for thyroid diseases and compared with findings of histopathologic examination. Time intensity curves for solid lesions were determined, excluding cases with secondary changes such as calcification, hemorrhage, necrosis and fibrosis. Three different patterns of time intensity curves were observed: rapid washout, delayed washout and no change. In our previous study, malignant grades of thyroid tumors were estimated immunohistochemically by epidermal growth factor receptor (EGFR) antibody. In most of malignant diseases and a few benign diseases that had marked cell proliferative activity with staining EGFR strongly, the time intensity curve displayed a delayed washout pattern, in which intensity was above 1/2- maximal value within 10 min after injection Gd-DTPA. Almost all benign diseases and a few well differentiated carcinomas displayed a rapid washout pastern, in which intensity was decreased to lower than 1/2 of peak grade within 10 min following injection and showed staining EGFR weakly. Benign diseases showing no change of time intensity curve, did not almost show aEGFR positive cell. These findings suggested that the time intensity curve obtained from dynamic MRI might indicate differentiated grades and cell proliferating activity of thyroid tumors.
  • Induction of manganese superoxide dismutase by thyroid stimulating hormone in rat thyroid cells, S Nishida, T Nakano, S Kimoto, T Kusunoki, K Suzuki, N Taniguchi, K Murata, TT Tomura, FEBS LETTERS, FEBS LETTERS, 416(1), 69 - 71, Oct. 1997 , Refereed
    Summary:Alterations in the superoxide dismutase (SOD) content of thyroid tissues occurring in association with thyroid dysfunction have been reported, In this study, the Mn-SOD content was found to increase in thyroid tissues of rats administered thyroid stimulating hormone (TSH) and in thyrocytes cultured in medium supplemented with TSH, Furthermore, in the thyroid glands of rats whose serum TSH level was elevated by inhibiting the synthesis of T3 and T4 by 6-methyl-2-thiouracil, the Mn-SOD increased as the TSH concentration increased, In the cultured thyrocytes, the increase in Mn-SOD induced by TSH was inhibited by the C-kinase inhibitor H7. These findings suggest the induction of Mn-SOD by TSH in thyroid cells and point to a role of C-kinase in this process, thereby indicating that a close relationship exists between the serum TSH level and the change in Mn-SOD content in thyrocytes with thyroid dysfunction, (C) 1997 Federation of European Biochemical Societies.
  • Study on cathepsin B and L in human thyroid tumors, T KUSUNOKI, S NISHIDA, K KABASHI, K MURATA, T TOMURA, PROTEASES INVOLVED IN CANCER, PROTEASES INVOLVED IN CANCER, 71 - 74, 1995 , Refereed
  • Study of type I and IV collagenase activity in human thyroid diseases, Takayuki Nakano, Takeshi Kusunoki, Kunihiko Funasaka, Kiyotaka Murata, Shozo Nishida, Takanori Tomura, Journal of Otolaryngology of Japan, Journal of Otolaryngology of Japan, 98(6), 937 - 9411073, 1995 , Refereed
    Summary:The activity of type I and IV collagenase was measured in thyroid tissue obtained from 6 non-diseased thyroids, 4 patients with Graves' diseases, 5 with follicular adenoma, 6 with papillary carcinoma and 4 with follicular carcinomas. The relationship between these enzyme activities and invasion or metastasis of the original tumors was studied. The activity of type I collagenase in papillary carcinomas and follicular carcinomas was higher than in non-diseased thyroids, Graves' disease and follicular adenoma. Carcinoma tissue with invasion beyond the capsule in particular had higher type I collagenase activity. Type IV collagenase activity in carcinoma with lymph node metastasis was higher than in non-diseased thyroids, Graves' disease and follicular adenoma, and especially higher than carcinoma without lymph node metastasis. These findings suggest that increased type I collagenase activity plays an important role in local invasion in thyroid carcinoma, and that increased type IV collagenase activity plays an important role in lymph node metastasis. © 1995, The Oto-Rhino-Laryngological Society of Japan, Inc. All rights reserved.
  • Study on Cathepsin B Activity in Human Thyroid Tumors, Takeshi Kusunoki, Shozo Nishida, Takayuki Nakano, Kunihiko Funasaka, Saori Kimoto, Kiyotaka Murata, Takanori Tomura, Auris Nasus Larynx, Auris Nasus Larynx, 22(1), 43 - 48, 1995 , Refereed
    Summary:Cathepsin B activity was measured in human thyroid tissue obtained surgically from 2 patients with Grave's disease, 3 with follicular adenoma, 4 with papillary carcinoma, and 4 with follicular carcinoma. Three normal thyroid tissues were also studied. Comparisons were made between cathepsin B activity and clinical findings, including histopathological diagnosis and the presence or absence of extra-capsular invasion and metastasis. The abilities of original tumors to degrade type I and type IV collagen were also measured. Mean cathepsin B activities of both specimens with benign and those with malignant disease were significantly higher than those of normal thyroid. On cases of thyroid carcinoma, those with extra-capsular invasion and metastasis had the highest cathepsin B activities. Cases with high cathepsin B activities also tended to show high type I and IV collagen degrading abilities. These findings suggest that cathepsin B plays a role in the development of extra-capsular invasion and lymph node metastasis in human thyroid tumors. © 1995, Society for Promotion of International Otorhinolaryngology (SPIO). All rights reserved.
  • Productivity of Thyroglobulin and Thyroid Hormone on Human Thyroid Cell in Collagen Gel Culture, Takeshi Kusunoki, Shozo Nishida, Masahiro Koezuka, Kiyotaka Murata, Takanori Tomura, Auris Nasus Larynx, Auris Nasus Larynx, 22(2), 120 - 127, 1995 , Refereed
    Summary:Thyroid cells obtained from 21 normal human thyroid tissue samples and 17 tissue samples from diseased thyroid, including one of Graves disease, 4 with follicular adenoma, 11 with papillary carcinoma, and 1 with follicular carcinoma, were cultured in collagen gel, and ability to produce thyroglobulin (Tg), triiodothyronine (T3) and thyroxine (T4) was determined. Changes in morphological characteristics and production of Tg, T3, and T4 induced by addition of thyroid stimulating hormone (TSH) to medium in collagen gel culture were also determined. Twenty of all cases exhibited positive reaction for Tg. No relationship was found between rate of positivity for Tg and pathologic diagnosis. Three with carcinoma showed positive reaction for T3, and 4 with carcinoma showed positive reaction for T4. Only for normal thyroid cells did addition of TSH to medium induce increase the percentage of colonies producing Tg or T4 and morphological changes including an enlarged follicular lumen and increase in the height of columnar epithelium. These findings suggest that thyroid cells in collagen culture develop in an in vivo-like fashion. In conclusion, collagen has important effects on cellular differentiation when included in extracellular matrix. © 1995, Society for Promotion of International Otorhinolaryngology (SPIO). All rights reserved.
  • Histopathological Findings of Human Thyroid Tumors and Signal Intensities of Magnetic Resonance Imaging (MRI), Takeshi Kusunoki, Kiyotaka Murata, Shozo Nishid, Takanori Tomura, Masaaki Inoue, Journal of Otolaryngology of Japan, Journal of Otolaryngology of Japan, 97(8), 1406 - 1411, 1994 , Refereed
  • CHANGES OF MYOCARDIAL METALLOTIONEIN ON ISOPROTERENOL-INDUCED MYOCARDIAL INJURY, K NAMIKAWA, Y OKAZAKI, S NISHIDA, S KIMOTO, F AKAI, T TOMURA, S HASHIMOTO, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 113(8), 591 - 595, Aug. 1993 , Refereed
    Summary:The amounts of myocardial metallothionein (MT) and heavy metal (Zn, Cu) levels during the early stage of the experimental myocardial infarction model induced by isoproterenol (Isp) administration were measured by an atomic absorption spectrophotometry. MT was measured by the Cd-hem method. Myocardial infarction was induced by the administration of 75 mg/kg i.p. of Isp to rats weighing 270 +/- 10 g. Thirty minutes after Isp injection, Zn and Cu levels began to decrease and 12 h later, reached the minimal values compared with the control value. The level of MT began to increase 3 h after the Isp injection and reached the maximal value at 12 h, although MT remained undetectable in the control myocardial tissue by the Cd-hem method. MT levels in the liver increased and total Zn and Cu were elevated compared with the control value 12 h after Isp administration. These results suggest that MT is produced in the myocardium after Isp administration, and that the roles of MT in the heart and the liver are different. It was thought that a rise in MT was induced for the protection of the myocardial cells to injury.
  • DRUG SENSITIVITY TEST FOR PRIMARY CULTURE OF HUMAN CANCER-CELLS USING COLLAGEN GEL EMBEDDED CULTURE AND IMAGE-ANALYSIS, M KOEZUKA, N KONDO, H KOBAYASHI, S HARA, M YASUTOMI, S NISHIDA, S HASHIMOTO, H ASANO, INTERNATIONAL JOURNAL OF ONCOLOGY, INTERNATIONAL JOURNAL OF ONCOLOGY, 2(6), 953 - 959, Jun. 1993 , Refereed
    Summary:A new drug sensitivity assay method using the collagen gel embedded culture and image analysis was developed. Human cancer cells cultured in collagen gels showed extremely high cloning efficiency and in vivo-like drug response. In addition, image analysis was used successfully for the first time to automatically discriminate cancer colonies from contaminating fibroblastic cells by taking advantage of the difference in shape between them after cultivation to determine the drug response of cultured cancer cells accurately and easily. A new approach for the practical use of the drug sensitivity test on human cancer cells is suggested.
  • MORPHOLOGICAL PROPERTIES OF HUMAN THYROID-TUMOR CELLS IN COLLAGEN GEL CULTURE AND METASTATIC OR INVASIVE ABILITY, S NISHIDA, K HOSOKAWA, T KUSUNOKI, M KOEZUKA, F AKAI, T NAKANO, K FUNASAKA, Y YOSHIOKA, K MURATA, S HASHIMOTO, TT TOMURA, HISTOLOGY AND HISTOPATHOLOGY, HISTOLOGY AND HISTOPATHOLOGY, 8(2), 329 - 337, Apr. 1993 , Refereed
    Summary:Using normal human thyroid cells and tumor cells, the reconstruction of various diseased cells in collagen gel as well as the relationship between the morphology of colonies in collagen-embedded culture and the biological behavior (benignity, malignancy, metastasis, and invasion) of the original tumors were studied. In collagen gel culture, normal thyroid cells reorganized follicle-like constructions, and follicular adenoma cells showed in vivo-like constructions. However, two different types of colonies were observed in cultures of cells from papillary carcinomas. One was the branching type with many outgrowths projecting to three dimensions and the other was the spherical type without any outgrowths. These spherical colonies were observed in all cases of papillary carcinoma, but varied from one case to another. Metastasis and invasion were detected during pathological examination in cases with a high ratio of spherical colonies. Our results indicate that cells from highly metastatic and invasive thyroid cancer form spherical colonies in the collagen gel culture, and that this collagen culture is a useful method for studying the heterogeneity of tumor cells as well as the metastasis and invasive ability of tumor cells in vitro.
  • MANGANESE SUPEROXIDE-DISMUTASE CONTENT AND LOCALIZATION IN HUMAN THYROID-TUMORS, S NISHIDA, F AKAI, H IWASAKI, K HOSOKAWA, T KUSUNOKI, K SUZUKI, N TANIGUCHI, S HASHIMOTO, TT TAMURA, JOURNAL OF PATHOLOGY, JOURNAL OF PATHOLOGY, 169(3), 341 - 345, Mar. 1993 , Refereed
    Summary:Manganese-containing superoxide dismutase (Mn-SOD) content and its immunohistochemical localization in human thyroid tumours and some other thyroid diseases were examined and compared with adjacent normal thyroid tissue. Enzyme-linked immunosorbent assay (ELISA) was used in this study for the measurement of Mn-SOD. The content of Mn-SOD tended to increase in diffuse hyperplasia, adenomatous goitre, and follicular adenoma. In papillary carcinoma, it was significantly higher than in adjacent normal thyroid tissue. Follicular carcinoma also revealed a markedly high Mn-SOD content. In the immunohistochemical study, adjacent normal thyroid tissue showed granular positive staining of Mn-SOD in the cytoplasm. An increase of Mn-SOD was observed in the papillary proliferative lesion of diffuse hyperplasia and in the follicles adjacent to lymphoid tissue in chronic thyroiditis with hypothyroidism. Strong positive staining of Mn-SOD was observed in papillary and follicular carcinomas, whereas in anaplastic carcinoma staining was markedly less intense. These results indicate that the Mn-SOD content varies according to the degree of differentiation of thyroid carcinomas.
  • Study on human diseased thyroid cells in collagen gel culture, Takeshi Kusunoki, Yasuhiro Yoshioka, Yosihiko Kohda, Kiyotaka Murata, Shozo Nishida, Takanori Tomura, Masahiro Koezuka, Fumihiko Ohta, Nippon Jibiinkoka Gakkai Kaiho, Nippon Jibiinkoka Gakkai Kaiho, 96(1), 35 - 40169, 1993 , Refereed
    Summary:Thyroid cells from 14 normal subjects, two patients with Grave's disease, four patients with follicular adenoma and eight patients with papillary carcinoma were cultured in collagen gel. The colonies of these cells were stereoscopically observed and their morphological characteristics were studied with regard to relation with pathological findings of mother tumor, extra-capsular invasion and metastatic potential. For normal thyroid, Grave's disease and follicular adenoma (except for one case), their own characteristic branching type colonies were found. For papillary carcinoma, both branching type and spheroid type of colonies were observed. The ratio of branching type/spheroid type varied individually in the patients with papillary carcinoma. However, the spheroid type was found to trend to be predominant in patients with extra-capsular invasion and/or lymph node metastasis. This means that the observation of spheroid type colonies in collagen gel is suggestive of risk of extra-capsular invasion or lymph node metastasis. From the obtained results, it seemed possible to diagnose poorly-differentiated cells in vitro by morphologically observing colonies of human thyroid papillary carcinoma cells developing in collagen gel. © 1993, The Oto-Rhino-Laryngological Society of Japan, Inc. All rights reserved.
  • Proliferating cell nuclear antigen (PCNA) on human diseased thyroid cells, Takeshi Kusunoki, Takayuki Nakano, Kunihiko Funasaka, Kiyotaka Murata, Shozou Nishida, Takanori Tomura, Shigeo Hashimoto, Fumihiko Ohta, Nippon Jibiinkoka Gakkai Kaiho, Nippon Jibiinkoka Gakkai Kaiho, 96(4), 651 - 658735, 1993 , Refereed
    Summary:Surgical materials obtained from 5 normal human thyroids and 19 diseased tissues (4 with Grave's disease, 4 adenomatous goiters, 3 follicular adenomas, 2 papillary carcinomas, 4 follicular carcinomas and 2 undifferentiated carcinomas) were investigated immunohistochemically using the avidin-biotin-peroxidase complex technique with anti-proliferating cell nuclear antigen (PCNA) antibody and epidermal growth factor receptor (EGFR) antibody. The results of this study suggested the following: (1) Labeling indexes of PCNA positive cell nuclei were shown in high grade malignant cases accompanied by capsular invasion and lymph node metastasis, in undifferentiated carcinoma cells, and in proliferative lesions of benign diseases. These results suggest that PCNA is indicative of proliferative ability in thyroid diseases. (2) On thyroid tumors, EGFR regions with strong positive staining showed many PCNA positive cell nuclei. EGF appears to have a significant association with proliferative ability in thyroid diseases. © 1993, The Oto-Rhino-Laryngological Society of Japan, Inc. All rights reserved.
  • STUDIES ON EARLY STAGE CHANGES OF PEROXIDE LIPID IN ISOPROTERENOL-INDUCED MYOCARDIAL INJURY, K NAMIKAWA, Y OKAZAKI, S NISHIDA, E KADOTA, T TOMURA, S HASHIMOTO, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 112(8), 557 - 562, Aug. 1992 , Refereed
    Summary:The amount of peroxide lipid in vivo in the early stage of the experimental model of myocardial infarction in a rat induced by the administration of isoproterenol (Isp) was measured as the value of malonic dialdehyde (MDA). The model of myocardial infarction was made by giving 75 mg/kg of Isp to the rat weighing 270 +/- 10 g. After the administration of Isp, the amounts of lipid in the serum and in the myocardial tissue were measured, and a blood chemistry test (glutamic oxaloacetic dehydrogenase, glutamic pyruvic transaminase, lactate dehydrogenase, free fatty acid, creatine kinase) was simultaneously carried out on the serum. The value of the amount of peroxide lipid in the serum began to elevate 3 h after the administration of Isp and reached a mximum value at 6 h. The value of the amount of peroxide lipid in the tissue began to elevate 30 min after the administration and reached a maximum at 3 h. Each blood chemistry disclosed the elevation 30 min after the administration. As mentioned above, the production of peroxide lipid in vivo on the myocardial disorder in the early stage after the administration of Isp and the biochemical changes showed a significant correlation. From these results it is suggested that the myocardial disorder induced by the administration of Isp has already developed at 30 min after the administration.
  • STUDIES ON EARLY CHANGES IN MYOCARDIAL ELECTROLYTES AND HISTOLOGICAL REACTION IN ISOPROTERENOL-INDUCED MYOCARDIAL INJURY, K NAMIKAWA, Y OKAZAKI, S NISHIDA, T TOMURA, S HASHIMOTO, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 111(4-5), 247 - 252, Apr. 1991 , Refereed
    Summary:To determine morphological changes in the very early stage of damage and recovery of myocardial fibers in relation to electrolytes and water content in the myocardium, experimental myocardial infarcts were produced after injection of 75 mg/kg isoproterenol (Isp) to rats weighing 270 +/- 10 g. Electrolytes were examined by an atomic absorption spectrophotometer, and histologic examination was performed by light and electron microscopy. At 30 min after Isp injection, Ca and water contents of myocardium increased, but Mg decreased. These myocardial damages showed a maximum level from 6 to 12 h after injection of Isp. It consisted of reversible injury of muscle fibers represented by eosinophilic and fatty degeneration, intermingled with coagulation necrosis of fibers which was irreversible. Most of these eosinophilic degeneration fibers recovered to normal fibers between 12 h and 3 d after the injection. The chronological changes of Ca, Mg, and water contents were well correlated to the morphological early changes of myocardial fibers, and this experimental model by using Isp may be useful for examining the recovery of damaged myocardial tissue and estimating the effects of therapeutics.
  • Morphological study of human thyroid papillary carinoma cells in collagen gel culture, Takeshi Kusunoki, Yasuhiro Yoshioka, Yoshihiko Kahda, Kiotaka Murata, Fumihiko Ohta, Shozou Nishida, Takanori Tomura, Masahiro Koezuka, Nippon Jibiinkoka Gakkai Kaiho, Nippon Jibiinkoka Gakkai Kaiho, 94(1), 16 - 21, 1991 , Refereed
    Summary:Human thyroid papillary carcinoma cells (surgical materials) were cultured in a monolayer or a collagen gel system. In the monolayer culture, degeneration and necrosis of cancer cells were observed after 10 days of culture. In the collagen gel culture, degenerative necrosis were observed 14 days culture. In histopathological observation monolayer cells proliferated in pavement-like form without forming any colony. On the contrary, collagen gel culture cells formed branching-type colonies and spheroid-type colonies. Those results suggested that papillary carcinoma cells can be cultured more like in vivo in collagen gel culture than monolayer culture. © 1991, The Oto-Rhino-Laryngological Society of Japan, Inc. All rights reserved.
  • IMMUNOHISTOCHEMICAL CHANGE OF ACTIN IN EXPERIMENTAL MYOCARDIAL-ISCHEMIA - ITS USEFULNESS TO DETECT VERY EARLY MYOCARDIAL DAMAGES, S NISHIDA, S HIRUMA, S HASHIMOTO, HISTOLOGY AND HISTOPATHOLOGY, HISTOLOGY AND HISTOPATHOLOGY, 2(4), 417 - 428, Oct. 1987 , Refereed
  • LECTIN (UEA-1) REACTION OF CAPILLARY ENDOTHELIUM WITH REFERENCE TO PERMEABILITY IN AUTOPSIED CASES OF CEREBRAL INFARCTION, E KADOTA, K TANJI, S NISHIDA, M TAKAHASHI, M MAEDA, S HIRUMA, Y ENOMOTO, S HASHIMOTO, F AKAI, HISTOLOGY AND HISTOPATHOLOGY, HISTOLOGY AND HISTOPATHOLOGY, 1(3), 219 - 226, Jul. 1986 , Refereed
  • IMMUNOHISTOCHEMICAL REACTION OF MYOCARDIAL FIBERS WITH ACTIN ANTISERUM IN AUTOPSY CASES OF MYOCARDIAL-INFARCTION, S HASHIMOTO, S NISHIDA, S HIRUMA, M TAKAHASHI, Y ENOMOTO, K SAKATANI, HISTOLOGY AND HISTOPATHOLOGY, HISTOLOGY AND HISTOPATHOLOGY, 1(3), 241 - 249, Jul. 1986 , Refereed
  • A MORPHOLOGICAL-STUDY ON THE EFFECTS OF COLLAGEN GEL MATRIX ON REGENERATION OF SEVERED RAT SCIATIC-NERVE IN SILICONE TUBES, T SATOU, S NISHIDA, S HIRUMA, K TANJI, M TAKAHASHI, S FUJITA, Y MIZUHARA, F AKAI, S HASHIMOTO, ACTA PATHOLOGICA JAPONICA, ACTA PATHOLOGICA JAPONICA, 36(2), 199 - 208, Feb. 1986 , Refereed
  • A MORPHOLOGICAL STUDY ON THE EFFECTS OF COLLAGEN GEL MATRIX ON REGENERATION OF SEVERED RAT SCIATIC NERVE IN SILICONE TUBES, Takao Satou, Shozo Nishida, Shingo Hiruma, Kurenai Tanji, Manabu Takahashi, Shuichiro Fujita, Yasuharu Mizuhara, Fumiharu Akai, Shigeo Hashimoto, Pathology International, Pathology International, 36(2), 199 - 208, 1986 , Refereed
    Summary:The present study is a chronological morphological examination on the effects of collagen gel matrix on regeneration of severed sciatic nerves. The nerves (5 mm length) were resected, and both the distal and proximal stumps were inserted into a silicone tube with 5 mm gap in between. In the test side, the gap in the tube was then injected with liquid collagen which gells in the tissue when reconstructed with a certain buffer solution. The gap space in the tube of the control side was left empty. In a chronological examination of the tissue in the tube, considerably more rapid growth of sprouting axons toward the distal stump in the test side was revealed in comparison with the control side. The cells, including both fibroblasts and larger Schwann cells, were less in number. More orderly directions were observed in the collagen matrix than in the control tube. The result indicates that regeneration of the peripheral nerves in the silicone tube can be improved, by using appropriate exogenous fine materials, collagen matrix. Copyright © 1986, Wiley Blackwell. All rights reserved
  • EXPERIMENTAL-STUDY OF WGA BINDING ON THE ENDOTHELIAL-CELL SURFACE IN CEREBRAL-ISCHEMIA, S NISHIDA, F AKAI, S HIRUMA, M MAEDA, K TANJI, S HASHIMOTO, HISTOLOGY AND HISTOPATHOLOGY, HISTOLOGY AND HISTOPATHOLOGY, 1(1), 69 - 74, Jan. 1986 , Refereed

Books etc

  • Alzheimer's disease pathogenesis-core concepts, shifting paradigms and therapeutic targets, Expression and cerebral function of amyloid precursor protein after rat traumatic brain injury, Joint author, Intech open access publisher,   2011 09
  • Study on cathepsin B and L in human thyroid tumors, Proteases involved in Cancer,   1995

Conference Activities & Talks

  • Macrophage inflammatory protein-1α induces osteoclast formation by activation of MEK/ERK pathway,   2010
  • Decreased Intestinal Detoxification Functions in an Absorption Process in Adjuvant-Induced Arthritis Rats,   2005
  • Enhancement of TNF-alpha induced antitumor activity by inhibition of C-kinase and its mechanism,   2004 10
  • Enhancement of TNF induced antitumor activity by C-kinase inhibitor,   2004 09
  • Apoptosis-inducing effect of mevastatin on hematopoietic tumor cell line,   2004 09
  • Enhancement of TNF-α induced antitumor activity by inhibition of C-kinase and its mechanism,   2003
  • Type IV collagen, type IV collagenase activity and ability of cell proliferation in human thyroid cancers,   2002 04
  • Change of map kinase signaling pathway in apoptotic cells induced by diethyldithiocarbamate (DDC),   2002
  • Apoptosis-inducing effect of a new bisphosphonate, YM529, on various hematopoietic of a new bisphosphonate, YM529, on various hematopoietic tumor cell lines,   2002
  • Type IV collagenase activity and immunostaining of type IV collagen in human thyroid tumors,   2001 11
  • Activity of type IV collagenase and immunostaining of type IV collagen in human thyroid,   2001 06
  • Proteases and their inhibitors in cholesteatoma,   2001 05
  • Immunostaining of type IV collagen, type IV collagenase and ability of cell proliferation in human thyroid cancers,   2000 12
  • Change of redox state induce apoptotic cell death without interfering with activation of Bcl-2 and HSP70,   2000 10
  • Mechanism of apoptosis regulated by redox state,   2000 10
  • Cathepsin L and its inhibitor in cheasteatoma of middle ear,   2000 05

Misc

  • 「地域チーム医療を担う薬剤師養成プログラム」における薬局実務実習の地域医療連携実習について 薬学部実習生の視点および評価から, 伊藤 麻祐, 藤本 美弥, 小川 充恵, 小枝 伸行, 山崎 肇, 篠原 裕子, 奥村 隆司, 中野 道雄, 安原 智久, 河野 武幸, 村岡 未彩, 西野 隆雄, 平田 收正, 井上 知美, 伊藤 栄次, 西田 升三, 小竹 武, 日本薬学会年会要旨集, 138年会, 4, 223, 223,   2018 03
  • 薬剤師を対象とした二次救命処置講習会の評価, 井上 知美, 小島 理恵, 岡田 ひとみ, 有元 秀樹, 窪田 愛恵, 平出 敦, 西田 升三, 石渡 俊二, 小竹 武, 日本薬学会年会要旨集, 138年会, 4, 226, 226,   2018 03
  • 心疾患患者における薬剤師の今後の役割と課題~フィジカルアセスメント~, 井上知美, 大鳥徹, 細見光一, 石渡俊二, 西田升三, 小竹武, 日本心臓病学会学術集会(Web), 65回, CS4, 2,   2017 09 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201702224726139292
  • 「地域チーム医療を担う薬剤師の養成プログラム」における薬薬学連携地域医療実務実習(八尾モデル) 薬学部実習生の視点から, 野村 加奈子, 中 雄一郎, 篠原 裕子, 奥村 隆司, 小川 充恵, 小枝 伸行, 山崎 肇, 井上 知美, 伊藤 栄次, 西田 升三, 小竹 武, 村岡 未彩, 西野 隆雄, 平田 收正, 日本薬学会年会要旨集, 137年会, 4, 221, 221,   2017 03
  • 「地域チーム医療を担う薬剤師育成プログラム」における地域連携医療関連実習について 薬学部実習生の視点から, 山下 佑麻, 假屋 幸音, 小川 充恵, 小枝 伸行, 山崎 肇, 篠原 裕子, 奥村 隆司, 中野 道雄, 村岡 未彩, 西野 隆雄, 平田 收正, 井上 知美, 伊藤 栄次, 西田 升三, 小竹 武, 日本薬学会年会要旨集, 137年会, 4, 221, 221,   2017 03
  • 「地域チーム医療を担う薬剤師養成プログラム」におけるチーム医療関連実習について 薬学部実習生の視点から, 假屋 幸音, 山下 佑麻, 小川 充恵, 小枝 伸行, 山崎 肇, 篠原 裕子, 奥村 隆司, 中野 道雄, 村岡 未彩, 西野 隆雄, 平田 收正, 井上 知美, 伊藤 栄次, 西田 升三, 小竹 武, 日本薬学会年会要旨集, 137年会, 4, 221, 221,   2017 03
  • Survey on Customer Satisfaction for Evaluation and Improvement of Physical Assessment Practical Training Seminar for Pharmacists, 大鳥徹, 井上知美, 細見光一, 中川博之, 高島敬子, 近藤尚美, 高田亜美, 伊藤栄次, 中山隆志, 和田哲幸, 石渡俊二, 前川智弘, 船上仁範, 中村真也, 窪田愛恵, 平出敦, 松山賢治, 西田升三, 社会薬学, 35, 2, 94‐101, 101,   2016 12 10 , 10.14925/jjsp.35.2_94, http://jglobal.jst.go.jp/public/201702220578908674
    Summary:<p>In the areas of home medical care and self-medication, the role of the pharmacist is growing, partly as a result of Japan's aging society and the need to reduce medical costs. In response, the Kinki University Faculty of Pharmacy implemented a physical assessment practical training seminar in order to improve the physical assessment skills of practicing pharmacists. A series of questionnaires were conducted among pharmacists to investigate their perceptions of physical assessment practical training seminars. The results of the questionnaires were analyzed using Customer Satisfaction (CS) analysis and text mining. Based on a 5-point scale (1-low∼5-high), questionnaires revealed satisfaction for physical assessment practical training seminars was 4.6±0.6 (Ave.±S.D.). CS analysis revealed that the items "lectures" and "case seminars" had the highest level of satisfaction. However, items showing low levels of satisfaction were "auscultation of respiratory sounds" and "SBAR (Situation, Background, Assessment, Recommendation)." Results of text mining suggested a relationship between "physical assessment" and "difficult". Analysis of the questionnaires showed a high level satisfaction with physical assessment practical training seminars, notably physical assessment practice methods. However, CS analysis and text mining indicate the finer techniques of physical assessment were difficult to acquire.</p>
  • CS分析(Customer Satisfaction analysis)による薬剤師のためのフィジカルアセスメント講習会の評価と改善, 大鳥 徹, 井上 知美, 細見 光一, 中川 博之, 高島 敬子, 近藤 尚美, 高田 亜美, 伊藤 栄次, 中山 隆志, 和田 哲幸, 石渡 俊二, 前川 智弘, 船上 仁範, 中村 真也, 窪田 愛恵, 平出 敦, 松山 賢治, 西田 升三, 社会薬学, 35, 2, 94, 101,   2016 12 , 10.14925/jjsp.35.2_94
    Summary:フィジカルアセスメント講習会に参加した143名を対象にアンケートを実施した。CS分析(Customer Satisfaction analysis)とテキストマイニングにより検討した。男性が39%、女性が61%と女性が多かった。勤務先は保険調剤薬局が38%、病院が62%となっており、病院勤務者が多かった。改善度は、数値が大きいほど改善の優先順位は高くなるが、改善度の値が大きい順に五つ項目を上げると、「心電図の読み方」「呼吸音の聴診」「SBAR」「時間は適当でしたか」「心音の聴診」であった。CSグラフにおいて、優先的に改善する必要がある項目がプロットされる「要改善項目」(第4象限)として抽出されてきたのは、「呼吸音の聴診」、「SBAR」であった。「心音・腸音の聴診」、「心電図の読み方」、「血圧測定」は評価が低く、総合評価への影響度も低い項目である「改善検討項目」として第3象限にプロットされた。「講義内容」、「症例に興味が持てたか」、「フィジカルアセスメントモデルの機能」、「時間は適当でしたか」は評点が高く、総合評価への影響度も高い項目である「重要維持項目」として第1象限にプロットされた。
  • 因子分析による薬剤師と薬学生のフィジカルアセスメントスキル修得に対する特性の比較検討, 大鳥 徹, 中川 博之, 高島 敬子, 近藤 尚美, 高田 亜美, 細見 光一, 井上 知美, 北小路 学, 西田 升三, 松山 賢治, 社会薬学, 35, Suppl., 46, 46,   2016 08
  • 肺がん患者に対するマグネシウム補充によるシスプラチン関連腎障害の抑制効果の検討, 木寺康裕, 川上尚人, 金田裕靖, 藤原季美子, 野村守弘, 千葉康敬, 西田升三, 山添譲, 鶴谷純司, 田村孝雄, 中川和彦, 日本肺癌学会総会号, 54, 5, 409, 409,   2014 10 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402211958618410
  • A search for the risk factors for hiccups and evaluation of antiemetic therapy in CDDP-based chemotherapy, using cluster analysis, Hajime Asano, Mizuki Watanabe, Akinori Kawaguchi, Masashi Yanae, Yoshinori Funakami, Tetsuyuki Wada, Sumio Matzno, Yuzuru Yamazoe, Shozo Nishida, Seiji Ichida, Japanese Journal of Cancer and Chemotherapy, 40, 8, 1031, 1036,   2013 08 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302231237895971
    Summary:Hiccups are often observed in patients treated with cisplatin (CDDP)-based chemotherapy. It has been reported that gender and specific dosages of CDDP and antiemetic drugs (e.g., dexamethasone and 5-HT3 receptor antagonist) using standard therapy are major risk factors in the onset of hiccups. Recently, aprepitant has been added to the antiemetic therapy in CDDP-based chemotherapy. However, it is not known how the onset of hiccups takes place in antiemetic therapy including aprepitant according to the guideline. In this study, we used cluster analysis to classify 229 patients treated with CDDP-based chemotherapy, to investigate the effect of antiemetic therapy on the onset of hiccups and chemotherapy-induced nausea and vomiting (CINV). Our analysis indicated that aprepitant was not a major risk factor for the onset of hiccups in the high CDDP dose group (≧70 mg/m2). However, an effect of antiemesis was confirmed in the standard therapy with aprepitant. In conclusion, we suggest that aprepitant is effective for CINV, without causing the onset of hiccups in patients treated with high-dose CDDP-based chemotherapy.
  • 在宅医・薬局間における連携実態調査報告, 伊藤武志, 川口範明, 古川諭, 柳江正嗣, 野村守弘, 椿正寛, 西田升三, 山添譲, 日本緩和医療学会学術大会プログラム・抄録集, 18回, 441, 441,   2013 06 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302281837678424
  • Invention of an Anti-hazard Stopper for Anti-cancer Drug Leakage Named MOREMASEM, ISHIWATA SHUNJI, TAGA ATSUSHI, FUJITA HIDEKI, SHINAGAWA TAKAYUKI, NISHIDA SHOZO, KITA AYAKO, SUGIURA REIKO, 医療薬学, 39, 5, 271, 275,   2013 05 , 10.5649/jjphcs.39.271, http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302260095590582
  • Bisphosphonatesおよびstatinsによる骨髄間質細胞でのOPG発現増強効果とその機序の解明, 椿正寛, 駒居真紀子, 小川直希, 山添譲, 向井淳治, 西田升三, 日本薬学会年会要旨集(CD-ROM), 133年会, 3, 208, 208,   2013 03 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302204765137470
  • Minodronateによる骨肉腫細胞でのRas経路阻害を介した転移抑制効果, 嶌岡弘高, 椿正寛, 駒居真紀子, 小川直希, 山添譲, 向井淳治, 西田升三, 日本薬学会年会要旨集(CD-ROM), 133年会, 4, 131, 131,   2013 03 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302264747555500
  • 多発性骨髄腫での抗がん剤多剤耐性獲得機序, 駒居真紀子, 椿正寛, 小川直希, 山添譲, 向井淳治, 西田升三, 日本薬学会年会要旨集(CD-ROM), 133年会, 4, 61, 61,   2013 03 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302278410424102
  • Study on Anti-Proliferative Activities of Cultured Cordyceps Militaris on Cancer Cells, HIGASHINO MASAYUKI, HORIE HIROSHI, ITO MASAKATSU, TANAKA KEIKO, FUKUDA YASUKI, NISHIDA SHOZO, MURATA KAZUYA, MATSUDA HIDEAKI, 日本補完代替医療学会誌, 10, 1, 51, 57,   2013 03 , 10.1625/jcam.10.51, http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302291179898143
  • 病院・在宅医・薬局間における薬剤提供情報の実態調査報告, 伊藤武志, 川口範明, 古川諭, 柳江正嗣, 野村守弘, 椿正寛, 西田升三, 山添譲, 日本在宅医療学会学術集会プログラム・抄録集, 24th, 107,   2013 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302204080147383
  • Evaluation of the Safety Precaution to Occupational Exposure to Antineoplastic Drugs in the Department of Pharmacy, NAKANO HIROYUKI, YOSHIDA JIN, KOMORI KEIKO, MOCHIZUKI CHIE, NAKATA YOKO, TEI GENSHIN, MASU YOSHIE, KODA SHIGEKI, NISHIDA SHOZO, 医療薬学, 39, 1, 1, 9,   2013 01 , 10.5649/jjphcs.39.1, http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302297592376870
  • シスプラチン後発品製剤の安全性に関する検討, 柳江正嗣, 椿正寛, 中尾真理子, 藤原季美子, 浅野肇, 森田哲也, 野村守弘, 山添譲, 西田升三, 日本医療薬学会年会講演要旨集, 22nd, 325,   2012 10 10 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201202216160771763
  • マグネシウム補充によるシスプラチン関連腎障害の抑制効果の検討, 木寺 康裕, 川上 尚人, 谷崎 潤子, 田中 薫, 上田 眞也, 岡本 渉, 仁科 慎一, 金田 裕靖, 藤原 季美子, 千葉 康敬, 西田 升三, 山添 譲, 倉田 宝保, 岡本 勇, 中川 和彦, 日本癌治療学会誌, 47, 3, 974, 974,   2012 10
  • Statinsによる骨肉腫細胞での血管新生促進因子抑制効果, 椿正寛, 柳江正嗣, 山添譲, 松岡寛, 西田升三, 日本薬学会年会要旨集, 132年会, 4, 238, 238,   2012 03 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201202245000002674
  • ゾレドロン酸投与状況調査と腎機能評価におけるeGFRの有用性の検討, 柳江正嗣, 中尾真理子, 藤原季美子, 椿正寛, 廣瀬雄司, 森田哲也, 山添譲, 西田升三, 日本医療薬学会年会講演要旨集, 21st, 238,   2011 09 09 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201102294230798161
  • オキシコドン徐放錠からフェンタニル貼付剤へのオピオイドローテーション時に影響する因子の調査, 森本 陽之, 川口 明範, 村岡 修, 市田 成志, 西田 升三, 森山 健三, 山添 譲, 日本薬学会年会要旨集, 131年会, 4, 300, 300,   2011 03
  • 支持療法変化に伴うm‐FOLFOX6治療患者に対する過敏症反応への影響, 木寺康裕, 佐藤太郎, 藤原季美子, 山添譲, 森山健三, 文田壮一, 上田眞也, 岡本渉, 岡本邦夫, 鶴谷純司, 宮崎昌樹, 岡本勇, 椿正寛, 西田升三, 中川和彦, 日本消化管学会総会学術集会プログラム・抄録集, 7th, 334,   2011 , http://jglobal.jst.go.jp/public/201102276174637488
  • 支持療法変化に伴うm‐FOLFOX6治療患者に対する過敏症反応への影響, 木寺康裕, 佐藤太郎, 藤原季美子, 山添譲, 森山健三, 文田壮一, 上田眞也, 岡本渉, 岡本邦夫, 鶴谷純司, 宮崎昌樹, 岡本勇, 椿正寛, 西田升三, 中川和彦, 日本臨床腫瘍学会学術集会プログラム・抄録集, 8th, 256,   2010 , http://jglobal.jst.go.jp/public/201002291438927260
  • 保険薬局および病院実務実習に対する学生の意識調査, 北小路学, 石渡俊二, 大床真美子, 船上仁範, 八軒浩子, 多賀淳, 八木秀樹, 和田哲幸, 田邊元三, 市田成志, 西田升三, 社会薬学, 25, 2, 34, 34,   2007 02 , http://jglobal.jst.go.jp/public/200902262219415130
  • Mevastatin induces Apoptosis in HL60 Cells, Nishida Shozo, Tsubaki Masanobu, Tanimori Yoshihiro, Bulletin of Pharmaceutical Research and Technology Institute, 13, 81, 90,   2005 03 05 , http://ci.nii.ac.jp/naid/40007000277
  • A New Bisphosphonate,YM529 induces Apoptosis in HL60 Cells Dependent on Decrease in Phosphorylated ERK, Nishida Shozo, Fujii Yoshiki, Yoshioka Shohei, Bulletin of Pharmaceutical Research and Technology Institute, 12, 107, 116,   2004 03 05 , http://ci.nii.ac.jp/naid/110001180548
  • C‐kinase阻害によるTNFα壊死効果の増強とその機序の解明, 西田升三, 椿正寛, 木下さおり, 吉岡昌平, 垣谷有紀, 藤井克樹, 戸村隆訓, 入交清博, 日本癌学会総会記事, 62回, 454, 454,   2003 08 , http://jglobal.jst.go.jp/public/200902284330163100
  • Apoptosis-inducing Effect of a New Bisphosphonate,YM529,on Various Hematopoietic Tumor Cell Lines, Nishida Shozo, Kikuichi Shigeru, Haga Hisae, Bulletin of Pharmaceutical Research and Technology Institute, 11, 95, 104,   2003 03 05 , http://ci.nii.ac.jp/naid/110000967596
  • Apoptosis in Palatine Tonsil Carcinomas, KUSUNOKI Takeshi, NISHIDA Shozo, OKITA Arihiro, TERAO Kyoichi, MIYAZAKI Katsuhiro, MURATA Kiyotaka, TOMURA Takanori, Practica Oto-Rhino-Laryngologica, 94, 6, 531, 535,   2001 06 , 10.5631/jibirin.94.531, http://ci.nii.ac.jp/naid/10008096768
    Summary:Apoptosis was observed in palatine tonsil carcinomas (squamous cell carcinoma; 11 cases) by TUNEL staining. The TUNEL labeling index (TLI) of carcinoma cells had no relation to TNM, staging, pathohistological differentiated grades or the efficiency of radiotherapy. TLI (means: 5.2%) of postradiotherapy cases was higher than that of preradiotherapy cases (1.9%). TUNEL positive cells were found in the tissues surrounding cancers. In the surrounding tissues, the TLI (means: 5.1%) of postradiotherapy cases was higher than that of preradiotherapy cases (2.0%). These findings indicated that apoptosis induced by radiotherapy could suppress cancer development. Reversely, the invasion and metastasis of surviving cancers may be promoted by apoptosis induced in the surrounding tissues.
  • Metalloproteinase-1 in human chronic otitis media, Takeshi Kusunoki, Shozo Nishida, Katsuhiro Miyazaki, Kyoichi Terao, Kiyotaka Murata, Takanori Tomura, Practica Otologica, 92, 11, 1255, 1258,   1999 11 , 10.5631/jibirin.92.1255, http://ci.nii.ac.jp/naid/10005266733
    Summary:We immunohistochemically examined expressions of metalloproteinase-1 (MMP-1) in both cholesteatoma and non-cholesteatoma granulation tissues. All 8 patients with cholesteatoma epithelia, 10 of 11 patients with cholesteatoma granulation, and 2 patients with cholesterin granuloma of 5 non-cholesteatoma patients exhibited expression of MMP-1. Histopathological examinations of these granulation tissues revealed proliferative fibroblast cells and many inflammatory cells with expression of MMP-1. Cholesteatoma subepithelial granulation (1 case) and noncholesteatoma granulation tissues without cholesterin granuloma (2 cases) did not exhibit expression of MMP-1. Collagen fibers occupied the granulative tissues on histopathological examinations. However, fibroblast and inflammatory cells were rarely found.
  • Cathepsin L and Cystatin a Expression in Chronic Otitis Media, Takeshi Kusunoki, Kazuo Kobashi, Shozo Nishida, Kiyotaka Murata, Takanori Tomura, Hiroaki Nakatani, Takaki Hiwasa, Practica Otologica, 92, 1, 101, 105,   1999 01 , 10.5631/jibirin.92.101, http://ci.nii.ac.jp/naid/10005032760
    Summary:We examined the cathepsin L activity and expression of cystatin A (an inhibitor of cathepsin L) in 7 cases of cholesteatoma epithelia, 3 cases of cholesteatoma subepithelial granulation, 3 cases of non-cholesteatoma granulation and 3 cases with an intact mucous membrane in the middle ear. The mean cathepsin L activity was higher in the diseased tissues than in the intact mucous membrane of the middle ear. Those granulation tissues with high cathepsin L activity resulted in extensive bony destruction in both cholesteatomas and non choleateatomas of the middle ear. Cystatin A expression was assessed by Western blotting. All of the cases with an intact mucous membrane in the middle ear exhibited no expression of cystatin A. 7 of the 10 cases with diseased tissues expressed cystatin A in their cholesteatoma epithelia, cholesteatoma subepitheial granulation or non cholesteatoma granulation tissues. No relationship between cystatin A expression and the level of cathepsin L activity could be observed. © 1999, The Society of Practical Otolaryngology. All rights reserved.
  • Enzyme activity and content of superoxide dismutase in SHRSP brain and myocardium, KIMOTO Saori, NISHIDA Shozo, FUNASAKA Kunihiko, NAKANO Takayuki, TERAMOTO Kazuhiro, TOMURA Takanori, Medical journal of Kinki University, 20, 2, 237, 242,   1995 12 , http://ci.nii.ac.jp/naid/110000061196
    Summary:The enzyme activity and content of manganese superoxide dismutase (Mn-SOD), an oxygen free-radical scavenger, were determined in the right and left myocardium and two specific brain regions of stroke-prone spontaneously hypertensive rats (SHRSP) and Wistar Kyoto rats (WKY) as controls. The Mn-SOD content in the SHRSP myocardium was higher than that in the WKY myocardium, but the Mn-SOD activity in the SHRSP myocardium was lower than that in the WKY myocardium. These findings indicate that enzymatically inactive or low-active Mn-SOD protein exists in the SHRSP myocardium. The Mn-SOD content and activity in the cortex and hippocampus of SHRSP was lower than that in the same regions of the WKY. These findings suggest that an oxygen free-radical is an important factor behind hypertensive diseases in the brain and myocardium.
  • Study of type I and IV collagenase activity in human thyroid diseases, Takayuki Nakano, Takeshi Kusunoki, Kunihiko Funasaka, Kiyotaka Murata, Shozo Nishida, Takanori Tomura, Journal of Otolaryngology of Japan, 98, 6, 937, 9411073,   1995 , 10.3950/jibiinkoka.98.937, http://ci.nii.ac.jp/naid/10006824529
    Summary:The activity of type I and IV collagenase was measured in thyroid tissue obtained from 6 non-diseased thyroids, 4 patients with Graves' diseases, 5 with follicular adenoma, 6 with papillary carcinoma and 4 with follicular carcinomas. The relationship between these enzyme activities and invasion or metastasis of the original tumors was studied. The activity of type I collagenase in papillary carcinomas and follicular carcinomas was higher than in non-diseased thyroids, Graves' disease and follicular adenoma. Carcinoma tissue with invasion beyond the capsule in particular had higher type I collagenase activity. Type IV collagenase activity in carcinoma with lymph node metastasis was higher than in non-diseased thyroids, Graves' disease and follicular adenoma, and especially higher than carcinoma without lymph node metastasis. These findings suggest that increased type I collagenase activity plays an important role in local invasion in thyroid carcinoma, and that increased type IV collagenase activity plays an important role in lymph node metastasis. © 1995, The Oto-Rhino-Laryngological Society of Japan, Inc. All rights reserved.
  • Change in superoxide dismutase activity in SHRSP brain, NISHIDA Shozo, KIMOTO Saori, FUNASAKA Kunihiko, NAKANO Takayuki, TERAMOTO Kazuhiro, KOBASHI Kazuo, KUSUNOKI Takeshi, TOMURA Takanori, Medical journal of Kinki University, 19, 4補冊, 47, 49,   1994 12 , http://ci.nii.ac.jp/naid/110000061157
    Summary:Clinical and experimental evidence demonstrates that nerve cells of spontaneously hypertensive rats (SHR) or stroke-prone spontaneously hypertensive rats (SHRSP) are more sensitive to ischemic injury that those of Wistar Kyoto rats (WKY), and that lipidperoxide increases significantly in the SHRSP brain with aging. To elucidate the vulnerability of SHRSP nerve cells, the activity of SOD, the enzyme that protects cells against reactive free radicals by catalyzing dismutation, was examined in the brain tissues (cerebral cortex and hippocampus)of SHRSP in comparison with that of WKY. There was no significant difference in cerebral cortices between 15-week-old SHRSP and WKY. However, at 31 week old close to the onset of stroke, overall SOD activities in the cerebral cortex of SHRSP were significantly lower than those in WKY. Moreover, SOD activity in the hippocampus, the region of selective neuronal vulnerability, was lower than that in the cerebral cortex of either SHRSP or WKY. These results indicate that low SOD activity in SHRSP is closely related to the vulnerability of SHRSP nerve cells and the increased lipidperoxide of SHRSP brain tissue.
  • Change of Cu, Zn-superoxide dismutase activity in SHRSP myocardium, NISHIDA Shozo, KIMOTO Saori, NAKANO Takayuki, TERAMOTO Kazuhiro, FUNASAKA Kunihiko, TOMURA Takanori, Medical journal of Kinki University, 18, 4補冊, 39, 41,   1993 12 , http://ci.nii.ac.jp/naid/110000061050
    Summary:Clinical and experimental evidence has demonstrated that the hypertrophied myocardial tissue is more sensitive to ischemic injury than the normal myocardium. To elucidate the vulnerability of SHRSP hypertrophied myocardium, the activity of Cu, Zn-SOD, an enzyme protecting the cells against reactive free radicals by catalyzing the dismutation, of the right and left ventricles in SHRSP was examined and compared with that in WKY. The Cu, Zn-SOD activity of the left ventricle in SHRSP was significantly lower than that in WKY. These findings indicate that the low Cu, Zn-SOD activity of SHRSP hypertrophic myocardium is closely related with the increased lipid peroxide content in the cell membrane of the SHRSP myocardium.
  • CHANGES OF MYOCARDIAL METALLOTIONEIN ON ISOPROTERENOL-INDUCED MYOCARDIAL INJURY, K NAMIKAWA, Y OKAZAKI, S NISHIDA, S KIMOTO, F AKAI, T TOMURA, S HASHIMOTO, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 113, 8, 591, 595,   1993 08 , 10.1248/yakushi1947.113.8_591, http://ci.nii.ac.jp/naid/110003649426
    Summary:The amounts of myocardial metallothionein (MT) and heavy metal (Zn, Cu) levels during the early stage of the experimental myocardial infarction model induced by isoproterenol (Isp) administration were measured by an atomic absorption spectrophotometry. MT was measured by the Cd-hem method. Myocardial infarction was induced by the administration of 75 mg/kg i.p. of Isp to rats weighing 270 +/- 10 g. Thirty minutes after Isp injection, Zn and Cu levels began to decrease and 12 h later, reached the minimal values compared with the control value. The level of MT began to increase 3 h after the Isp injection and reached the maximal value at 12 h, although MT remained undetectable in the control myocardial tissue by the Cd-hem method. MT levels in the liver increased and total Zn and Cu were elevated compared with the control value 12 h after Isp administration. These results suggest that MT is produced in the myocardium after Isp administration, and that the roles of MT in the heart and the liver are different. It was thought that a rise in MT was induced for the protection of the myocardial cells to injury.
  • Change of superoxide dismutase content in SHRSP myocardium, NISHIDA Shozo, KIMOTO Saori, NAKANO Takayuki, TERAMOTO Kazuhiro, FUNASAKA Kunihiko, SUZUKI Keiichirou, TANIGUCHi Naoyuki, TOMURA Takanori, Medical journal of Kinki University, 17, 4補冊, 35, 37,   1992 12 , http://ci.nii.ac.jp/naid/110000060936
    Summary:To elucidate the vulunerable myocardium of SHRSP, the content of Mn-SOD, an enzyme protecting the cell from reactive free radicals by catalyzing the dismutation, of right or left ventricles in SHRSP, were examined and compared with that in WKY. The Mn-SOD content in the left ventricle in the SHRSP was significantly higher than that in the WKY. Moreover, in SHRSP, the Mn-SOD content in the left ventricle was significantly higher than that in the right ventricle. These findings suggest that the high Mn-SOD content of SHRSP hypertrophied myocardium is due to the increased oxygen stress in the mitochondria.
  • STUDIES ON EARLY STAGE CHANGES OF PEROXIDE LIPID IN ISOPROTERENOL-INDUCED MYOCARDIAL INJURY, K NAMIKAWA, Y OKAZAKI, S NISHIDA, E KADOTA, T TOMURA, S HASHIMOTO, YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN, 112, 8, 557, 562,   1992 08 , 10.1248/yakushi1947.112.8_557, http://ci.nii.ac.jp/naid/110003649346
    Summary:The amount of peroxide lipid in vivo in the early stage of the experimental model of myocardial infarction in a rat induced by the administration of isoproterenol (Isp) was measured as the value of malonic dialdehyde (MDA). The model of myocardial infarction was made by giving 75 mg/kg of Isp to the rat weighing 270 +/- 10 g. After the administration of Isp, the amounts of lipid in the serum and in the myocardial tissue were measured, and a blood chemistry test (glutamic oxaloacetic dehydrogenase, glutamic pyruvic transaminase, lactate dehydrogenase, free fatty acid, creatine kinase) was simultaneously carried out on the serum. The value of the amount of peroxide lipid in the serum began to elevate 3 h after the administration of Isp and reached a mximum value at 6 h. The value of the amount of peroxide lipid in the tissue began to elevate 30 min after the administration and reached a maximum at 3 h. Each blood chemistry disclosed the elevation 30 min after the administration. As mentioned above, the production of peroxide lipid in vivo on the myocardial disorder in the early stage after the administration of Isp and the biochemical changes showed a significant correlation. From these results it is suggested that the myocardial disorder induced by the administration of Isp has already developed at 30 min after the administration.
  • Studies on early changes in myocardial electrolytes and histological reaction in isoproterenol-induced myocardial injury, K. Namikawa, Y. Okazaki, S. Nishida, T. Tomura, S. Hashimoto, Yakugaku Zasshi, 111, 4〜5, 247, 252,   1991 05 , 10.1248/yakushi1947.111.4-5_247, http://ci.nii.ac.jp/naid/110003649226
    Summary:To determine morphological changes in the very early stage of damage and recovery of myocardial fibers in relation to electrolytes and water content in the myocardium, experimental myocardial infarcts were produced after injection of 75 mg/kg isoproterenol (Isp) to rats weighing 270±10g. Electrolytes were exmined by an atomic absorption spectrophotometer, and histologic examination was performed by light and electron microscopy. At 30 min after Isp injection, Ca and water contents of myocardium increased, but Mg decreased. These myocardial damages showed a maximum level from 6 to 12 h after injection of Isp. It consisted of reversible injury of muscle fibers represented by eosinophilic and fatty degeneration, intermingled with coagulation necrosis of fibers which was irreversible. Most of these eosinophilic degeneration fibers recovered to normal fibers between 12 h and 3 d after the injection. The chronological changes of Ca, Mg, and water contents were well correlated to the morphological early changes of myocardial fibers, and this experimental model by using Isp may be useful for examining the recovery of damaged myocardial tissue and estimating the effects of therapeutics.
  • DNA polymorphism and protein analysis on SDS-PAGE in SHR, OH'HARA Ikuko, MATSUOKA Hiroki, TAKADA Kazuo, SAWAGUCHI Hirochiyo, KAWASHIMA Akira, NISHIDA Shozo, TOMURA Takanori, Medical journal of Kinki University, 15, 4, 15, 18,   1990 12 30 , http://ci.nii.ac.jp/naid/110000060712
    Summary:The double digestion of DNA from spontaneously hypertensive rat (SHR) liver with HindIII/PvuII, revealed an extra band on 0.8% agarose gel electrophoresis but not that of DNA from Wistar-Kyoto rats (WKY). Protein analysis of kidneys by SDS-PAGE also showed different patterns between SHR and WKY.
  • DNA repair capacity in SHRSP, NISHIDA Shozo, OH'HARA Ikuko, TOMURA Takanori, Medical journal of Kinki University, 14, 4補冊, 13, 17,   1989 12 , http://ci.nii.ac.jp/naid/110000060593
    Summary:The present study was undertaken to extract crude enzymes from the liver of stroke-prone spontaneously hypertensive rats (SHRSP) and Wistar-Kyoto rats (WKY), and to compare repair endonuclease and DNA polymerase activity between SHRSP and WKY. Repair endonuclease activity was lower in SHRSP than in WKY (p<0.05). DNA polymerase activity was also low in SHRSP (p<0.01 ). The decrease in DNA repair capacity in SHRSP may be related to aging and also to hypertension and stroke.
  • An experimental pathomorphological study on recovery process from isoproterenol induced myocardial injury in special reference to changes of electrolytes, ENOMOTO Yoshiki, HASHIMOTO Shigeo, NISHIDA Shozo, NAMIKAWA Kiyohiro, Medical journal of Kinki University, 13, 4,   1988 12 25 , http://ci.nii.ac.jp/naid/110000060309
  • Preparation and use in immunohistology of antibody for Type IV collagen, KOEZUKA Masahiro, TAKAHASHI Hitoshi, NISHIDA Shozo, HIRUMA Shingo, HASHIMOTO Shigeo, Medical journal of Kinki University, 11, 2, 329, 330,   1986 06 25 , http://ci.nii.ac.jp/naid/110000059990
  • Production of the monoclonal antibody against chicken gizzard muscle actin and chicken skeletal muscle actin by HVJ M protein, HIRUMA Shingo, NISHIDA Shozo, HASHIMOTO Shigeo, WATANABE Masayasu, WADA Takuya, Medical journal of Kinki University, 10, 4, 453, 454,   1985 12 25 , http://ci.nii.ac.jp/naid/110000059915
  • Reactivity of anti-actin antibody in human and experimental myocardial infarction, NISHIDA Shozo, HIRUMA Shingo, MAEKURA Toshiharu, SAKAGUCHI Masatomo, HASHIMOTO Shigeo, Medical journal of Kinki University, 10, 4, 501, 502,   1985 12 , http://ci.nii.ac.jp/naid/110000059939
  • Study of Peripheral Nerve Regeneration-(1) : Effect of Collagen Matrix, SATOU Takao, HIRUMA Shingo, NISHIDA Shozo, TAMURA Yoshisada, ENOMOTO Yoshiki, MIZUHARA Yasuharu, TANJI Kurenai, HASHIMOTO Shigeo, Medical journal of Kinki University, 9, 4, 607, 608,   1984 12 30 , http://ci.nii.ac.jp/naid/110000060453
  • Rho/YAP経路活性化を介したRHAMM及びCXCR4過剰発現は転移を促進する(Overexpression of RHAMM and CXCR4 via activation of Rho/YAP pathway promotes tumor metastasis), 源野 秀次, 椿 正寛, 武田 朋也, 田畑 光希, 西田 升三, 日本癌学会総会記事, 78回, P, 2052,   2019 09
  • 骨髄腫分泌HGFは骨芽細胞におけるRANKL発現増加を介して骨破壊を促進する(Myeloma cellssecreted HGF accelerate the bone destruction via increased RANKL expression in osteoblasts), 田畑 光希, 椿 正寛, 武田 朋也, 関 しおり, 西田 升三, 日本癌学会総会記事, 78回, P, 2088,   2019 09
  • MEK阻害薬は抗がん剤誘発末梢神経障害を抑制できる(MEK inhibitor suppressed the anticancer drug-induced neuropathy), 加藤 菜月, 椿 正寛, 武田 朋也, 立石 敬典, 西田 升三, 日本癌学会総会記事, 78回, P, 3143,   2019 09
  • シンバスタチンによるオキサリプラチン誘発末梢神経障害抑制及び抗腫瘍作用増強効果(Simvastatin inhibited the oxaliplatin-induced neuropathy and potentiate antitumor effect of oxaliplatin), 立石 敬典, 椿 正寛, 武田 朋也, 加藤 菜月, 西田 升三, 日本癌学会総会記事, 78回, P, 3176,   2019 09
  • ベムラフェニブによるBRAF変異大腸癌でのオキサリプラチン及び5-FU殺細胞作用増強効果(Vemurafenib enhances sensitivity of oxaliplatin and 5-fluorourasil in BRAF-mutated colorectal cancer), 地主 みなみ, 椿 正寛, 武田 朋也, 関 しおり, 西田 升三, 日本癌学会総会記事, 78回, P, 3154,   2019 09
  • 大腸がんにおけるPI3K/Akt経路活性化を介したMEK阻害薬耐性機構(Activation of PI3K/Akt pathway contributes MEK inhibitor resistance in colon cancer), 椿 正寛, 武田 朋也, 加藤 菜月, 立石 敬典, 西田 升三, 日本癌学会総会記事, 78回, P, 3156,   2019 09
  • シグナル活性化を介したHIF-1過剰発現が骨髄腫でのメルファラン耐性に関与する(Overexpression of HIF-1alpha by activation of signal pathway involved with melphalan resistance in myeloma), 松田 拓弥, 椿 正寛, 武田 朋也, 田畑 光希, 関 しおり, 西田 升三, 日本癌学会総会記事, 78回, P, 3163,   2019 09
  • ダカルバジン及びスタチン併用療法は転移における生存率を改善できる(Combination treatment with dacarbazine and statins improved survival rate in melanoma metastasis-bearing mice), 西田 升三, 椿 正寛, 武田 朋也, 地主 みなみ, 関 しおり, 日本癌学会総会記事, 78回, P, 3338,   2019 09
  • ソラフェニブによる悪性黒色腫での受容体型チロシンキナーゼ抑制を介した腫瘍増殖・転移抑制効果(Sorafenib suppresses the tumor growth and metastasis of melanoma through suppression of receptor tyrosine kinase pathway), 武田 朋也, 椿 正寛, 地主 みなみ, 源野 秀次, 西田 升三, 日本癌学会総会記事, 78回, P, 3339,   2019 09
  • ERK1/2, Akt, NF-κB活性化を介したBim発現低下が骨髄腫での多剤耐性に中心的役割を果たす(Decreasing Bim expression via activated ERK, Akt, and NF-kappaB play a central role in multidrugresistant myeloma cells), 関 しおり, 椿 正寛, 武田 朋也, 地主 みなみ, 西田 升三, 日本癌学会総会記事, 78回, P, 3358,   2019 09
  • Dimethyl fumarateによるNF-kappaB抑制を介した腫瘍増殖及び転移抑制効果(Dimethyl fumarate suppresses the tumor growth and metastasis through suppression of NF-kappaB), 武田 朋也, 椿 正寛, 浅野 良太, 川島 啓司, 田畑 光希, 西田 升三, 日本癌学会総会記事, 77回, 389, 389,   2018 09
  • レバミピドによる口腔粘膜細胞でのAkt/mTOR経路活性化を介した抗がん剤誘導細胞死抑制効果(Rebamipide suppressed anticancer drugs-induced cell death via Akt/mTOR activation in oral mucosal keratinocytes), 川島 啓司, 椿 正寛, 武田 朋也, 浅野 良太, 藤本 伸一郎, 西田 升三, 日本癌学会総会記事, 77回, 694, 694,   2018 09
  • シグナル伝達因子活性化によるBim発現低下を介したアドリアマイシン耐性獲得機序(Downregulation of Bim via activation of signal molecules plays a central role in adriamycin resistant-myeloma cells), 河本 雄一, 椿 正寛, 武田 朋也, 浅野 良太, 西田 升三, 日本癌学会総会記事, 77回, 834, 834,   2018 09
  • PKC阻害剤による抗がん剤誘発末梢神経障害抑制効果(PKC inhibitor suppressed the anticancer drug-induced neuropathy), 加藤 菜月, 椿 正寛, 武田 朋也, 河本 雄一, 川島 啓司, 西田 升三, 日本癌学会総会記事, 77回, 854, 854,   2018 09
  • スタチンによる造血器腫瘍でのRas経路阻害を介したアポトーシス誘導効果(Statins induce apoptosis via inhibition of Ras/ERK and Ras/mTOR signaling pathways in hematopoietic tumor), 西田 升三, 椿 正寛, 武田 朋也, 藤原 大一朗, 河本 雄一, 藤本 伸一郎, 日本癌学会総会記事, 77回, 1460, 1460,   2018 09
  • Statins及びダカルバジン併用投与による腫瘍増殖及び転移抑制効果(Combined treatment of statins and dacarbazine inhibit tumor growth and metastasis in melanoma), 椿 正寛, 武田 朋也, 川島 啓司, 地主 みなみ, 西田 升三, 日本癌学会総会記事, 77回, 1605, 1605,   2018 09
  • 新規NF-kappaB/STAT3阻害剤による多発性骨髄腫でのアポトーシス誘導効果(A novel NF-kappaB/STAT3 inhibitor, bavachin, induces apoptosis in multiple myeloma cell lines), 浅野 良太, 椿 正寛, 武田 朋也, 加藤 菜月, 田畑 光希, 西田 升三, 日本癌学会総会記事, 77回, 1709, 1709,   2018 09
  • HGFは骨髄間質細胞及び骨芽細胞におけるRANKL発現を増加させる(HGF accelerate RANKL expression in bone marrow stromal cells and osteoblasts), 田畑 光希, 椿 正寛, 武田 朋也, 加藤 菜月, 西田 升三, 日本癌学会総会記事, 77回, 2151, 2151,   2018 09
  • 多発性骨髄腫でのメルファラン耐性にシグナル伝達経路活性化によるHIF-1alpha過剰発現が関与する, 友成 佳加, 椿 正寛, 武田 朋也, 藤本 伸一郎, 西田 升三, 日本癌学会総会記事, 76回, P, 1367,   2017 09
  • Src阻害はRANK/RANKLによるCAM-DRを克服する, 西田 升三, 椿 正寛, 武田 朋也, 友成 佳加, 川島 啓司, 日本癌学会総会記事, 76回, P, 1369,   2017 09
  • Rebamipideは口腔癌において5-fluorourasil感受性を増強させる, 川島 啓司, 椿 正寛, 武田 朋也, 河本 雄一, 西田 升三, 日本癌学会総会記事, 76回, P, 1378,   2017 09
  • CMLにおけるイマチニブ耐性にMET/ERK及びMET/JNK経路が関与する, 椿 正寛, 武田 朋也, 河本 雄一, 藤本 伸一郎, 西田 升三, 日本癌学会総会記事, 76回, E, 2036,   2017 09
  • 新規NIK阻害剤Mangiferinによる転移・腫瘍増殖抑制効果, 武田 朋也, 椿 正寛, 浅野 良太, 川島 啓司, 西田 升三, 日本癌学会総会記事, 76回, J, 2107,   2017 09
  • PioglitazoneはSTAT3阻害を介してSurvivinの発現低下及びAIFの発現増加によりアポトーシスを誘導する, 浅野 良太, 椿 正寛, 武田 朋也, 河本 雄一, 西田 升三, 日本癌学会総会記事, 76回, P, 2347,   2017 09
  • MIP-1alphaオートクラインは多発性骨髄腫でのメルファラン感受性を低下させる, 河本 雄一, 椿 正寛, 武田 朋也, 友成 佳加, 眞下 恵次, 阪口 勝彦, 西田 升三, 日本癌学会総会記事, 76回, J, 3113,   2017 09
  • 新規NIK阻害薬によるNF-kappaB経路抑制を介したアポトーシス誘導機序, 武田 朋也, 椿 正寛, 木野 稔己, 友成 佳加, 眞下 恵次, 阪口 勝彦, 西田 升三, 日本癌学会総会記事, 75回, J, 2032,   2016 10
  • 慢性骨髄性白血病におけるMET活性化はイマチニブ耐性に寄与する, 椿 正寛, 武田 朋也, 木野 稔己, 友成 佳加, 藤本 伸一郎, 西田 升三, 日本癌学会総会記事, 75回, J, 2035,   2016 10
  • HIF-1alphaの過剰発現は多発性骨髄腫でのメルファラン耐性に寄与する, 西田 升三, 椿 正寛, 武田 朋也, 木野 稔己, 友成 佳加, 藤本 伸一郎, 日本癌学会総会記事, 75回, J, 3021,   2016 10
  • RANK/RANKLによる細胞接着を介した抗がん剤耐性機構の解析, 木野 稔己, 椿 正寛, 武田 朋也, 友成 佳加, 眞下 恵次, 藤原 大一朗, 阪口 勝彦, 西田 升三, 日本癌学会総会記事, 75回, P, 3262,   2016 10
  • Mangiferinは多発性骨髄腫において抗がん剤感受性を増強させる, 友成 佳加, 椿 正寛, 武田 朋也, 木野 稔己, 藤原 大一朗, 阪口 勝彦, 西田 升三, 日本癌学会総会記事, 75回, P, 3285,   2016 10
  • Dimethyl fumarateによるNF-kappaB阻害を介したアポトーシス誘導効果, 木野 稔己, 椿 正寛, 武田 朋也, 小川 直希, 石坂 敏彦, 西田 升三, 日本癌学会総会記事, 74回, P, 1189,   2015 10
  • PKC阻害剤はオキサリプラチン誘導末梢神経障害を抑制するとともに抗腫瘍効果を増強させる, 椿 正寛, 武田 朋也, 中橋 拓也, 小川 直希, 石坂 敏彦, 西田 升三, 日本癌学会総会記事, 74回, J, 1192,   2015 10
  • Bisphosphonates及びstatinsによる骨髄腫でのRas経路阻害を介したMIP-1alpha発現抑制効果, 武田 朋也, 椿 正寛, 木野 稔己, 眞下 恵次, 藤原 大一朗, 阪口 勝彦, 西田 升三, 日本癌学会総会記事, 74回, J, 1209,   2015 10
  • Survivinの発現増加が多発性骨髄腫におけるvincristine耐性に中心的な役割を果たす, 西田 升三, 椿 正寛, 武田 朋也, 眞下 恵次, 藤原 大一朗, 阪口 勝彦, 日本癌学会総会記事, 74回, P, 2314,   2015 10
  • RANKL/RANKシステムによる乳がんでのNF-kappaB活性化を介したEMT促進効果(Activation of NF-kappaB by the RANKL/RANK system induces EMT in mammary tumor cell lines), 武田 朋也, 椿 正寛, 嶌岡 弘高, 坂本 洸太郎, 藤田 亜梨沙, 小川 直希, 山添 譲, 向井 淳治, 西田 升三, 日本癌学会総会記事, 73回, J, 2076,   2014 09
  • StatinsによるRho経路阻害を介した腫瘍増殖・転移抑制効果(Statins inhibit tumor growth and metastasis through the suppression of Rho signaling pathway), 坂本 洸太郎, 椿 正寛, 武田 朋也, 嶌岡 弘高, 藤田 亜梨沙, 眞下 恵次, 藤原 大一朗, 阪口 勝彦, 西田 升三, 日本癌学会総会記事, 73回, J, 3050,   2014 09
  • RANK/RANKLシステムによるシグナル伝達経路活性化を介した多剤耐性獲得機序(Induction of multidrug resistance in multiple myeloma by RANK/RANKL system mediates activation of signaling pathway), 椿 正寛, 武田 朋也, 坂本 洸太郎, 嶌岡 弘高, 藤田 亜梨沙, 眞下 恵次, 藤原 大一朗, 阪口 勝彦, 西田 升三, 日本癌学会総会記事, 73回, J, 1112,   2014 09
  • Src阻害に基づく抗がん剤耐性多発性骨髄腫での耐性克服効果(By inhibiting Src overcome multidrug resistance in multidrug-resistant myeloma cells), 藤田 亜梨沙, 椿 正寛, 武田 朋也, 嶌岡 弘高, 坂本 洸太郎, 小川 直希, 山添 譲, 向井 淳治, 西田 升三, 日本癌学会総会記事, 73回, J, 1114,   2014 09
  • ビスフォスフォネート及びスタチンによるMIP-1alpha分泌抑制効果(Bisphosphonates and statin inhibits MIP-1alpha secretion in myeloma cells), 西田 升三, 椿 正寛, 武田 朋也, 嶌岡 弘高, 坂本 洸太郎, 藤田 亜梨沙, 小川 直希, 山添 譲, 向井 淳治, 日本癌学会総会記事, 73回, P, 2239,   2014 09
  • 大腸癌でのRas経路阻害による抗がん剤感受性増強効果(Inhibition of Ras pathway enhances the cytotoxic effects of anti-cancer drug in colon cancer), 嶌岡 弘高, 椿 正寛, 武田 朋也, 坂本 洸太郎, 藤田 亜梨沙, 眞下 恵次, 藤原 大一朗, 阪口 勝彦, 西田 升三, 日本癌学会総会記事, 73回, P, 1387,   2014 09
  • BisphosphonatesによるRas阻害を介した造血器腫瘍でのアポトーシス誘導機序(Nitrogen-containing bisphosphonates induce apoptosis of hematopoietic tumor cells via inhibition of Ras pathways), 嶌岡 弘高, 椿 正寛, 駒居 真紀子, 坂本 洸太郎, 小川 直希, 眞下 恵次, 藤原 大一朗, 山添 譲, 阪口 勝彦, 向井 淳治, 西田 升三, 日本癌学会総会記事, 72回, 191, 191,   2013 10
  • RANK/RANKLを介した抗がん剤耐性獲得機序(Multi-drug resistance in multiple myeloma mediates RANK/RANKL system), 椿 正寛, 駒居 真紀子, 嶌岡 弘高, 坂本 洸太郎, 小川 直希, 眞下 恵次, 藤原 大一朗, 山添 譲, 阪口 勝彦, 向井 淳治, 西田 升三, 日本癌学会総会記事, 72回, 192, 192,   2013 10
  • シグナル伝達因子活性化による多発性骨髄腫での抗がん剤耐性獲得機序(Multidrug-resistance in multiple myeloma mediates activation of signal transduction molecules), 駒居 真紀子, 椿 正寛, 嶌岡 弘高, 坂本 洸太郎, 小川 直希, 眞下 恵次, 藤原 大一朗, 山添 譲, 阪口 勝彦, 向井 淳治, 西田 升三, 日本癌学会総会記事, 72回, 467, 467,   2013 10
  • TNF-alphaオートクライン阻害による抗がん剤殺細胞作用増強効果(Inhibition of the tumor necrosis factor-alpha autocrine enhances the sensitivity of myeloma cells to anticancer drugs), 西田 升三, 椿 正寛, 駒居 真紀子, 嶌岡 弘高, 坂本 洸太郎, 小川 直希, 眞下 恵次, 藤原 大一朗, 山添 譲, 阪口 勝彦, 向井 淳治, 日本癌学会総会記事, 72回, 470, 470,   2013 10
  • StatinsによるRho/LINK経路阻害を介した転移抑制効果(Statins inhibit metastasis through the suppression of Rho/LIMK pathway), 坂本 洸太郎, 椿 正寛, 駒居 真紀子, 嶌岡 弘高, 小川 直希, 眞下 恵次, 藤原 大一朗, 山添 譲, 阪口 勝彦, 向井 淳治, 西田 升三, 日本癌学会総会記事, 72回, 495, 495,   2013 10
  • 多発性骨髄腫でのMDR1、survivinの過剰発現及びBimの発現低下を介した抗がん剤耐性獲得(Overexpression of MDR1 and survivin, and decreased Bim expression mediate multidrug-resistance in multiple myeloma), 駒居 真紀子, 椿 正寛, 小川 直希, 山添 譲, 向井 淳治, 西田 升三, 日本癌学会総会記事, 71回, 326, 326,   2012 08
  • ミノドロネートによる骨肉腫細胞での転移抑制効果(Reduction of metastasis in osteosarcoma by minodronic acid), 西田 升三, 椿 正寛, 駒井 真紀子, 小川 直希, 山添 譲, 向井 淳治, 日本癌学会総会記事, 71回, 504, 504,   2012 08
  • Pioglitazoneによる抗がん剤殺細胞増強効果(Pioglitazone enhances the cytotoxic effects of anti-cancer drugs in head and neck carcinoma), 椿 正寛, 駒居 真紀子, 小川 直希, 山添 譲, 向井 淳治, 西田 升三, 日本癌学会総会記事, 71回, 569, 569,   2012 08
  • YM529/ONO-5920による悪性黒色腫細胞での肺転移抑制効果(Nitrogen-containing bisphosphonate, YM529/ONO-5920, inhibits tumor metastasis in mouse melanoma), 西田 升三, 椿 正寛, 柳江 正嗣, 木寺 康裕, 尾垣 光彦, 日本癌学会総会記事, 70回, 121, 121,   2011 09
  • Statinによる血管新生促進因子発現阻害効果(Blockade of the Ras/MEK/ERK and Ras/PI3K/Akt pathways by statins reduces angiogenic factors in mouse osteosarcoma), 椿 正寛, 柳江 正嗣, 木寺 康裕, 尾垣 光彦, 西田 升三, 日本癌学会総会記事, 70回, 125, 125,   2011 09
  • NF-κB阻害剤はマトリックスメタロプロテイナーゼの発現および活性を阻害することにより腫瘍細胞浸潤および転移を抑制する(NF-kappaB inhibitor suppresses tumor cell invasion and metastasis by inhibiting the expression and activities of matrix metalloproteinases), 金子 淳一, 椿 正寛, 山添 譲, 谷森 佳弘, 木寺 康裕, 柳江 正嗣, 西田 升三, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 4P, 0992,   2010 12
  • PREVENTIVE APPROACH TO HYPERSENSITIVITY REACTION INDUCED BY OXALIPLATIN, Kidera Yasuhiro, Satoh Taroh, Fujiwara Kimiko, Yamazoe Yuzuru, Moriyama Kenzo, Fumita Soichi, Ueda Shinya, Okamoto Wataru, Okamoto Kunio, Tsurutani Junji, Miyazaki Masaki, Okamoto Isamu, Tsubaki Masahiro, Nishida Shozo, Nakagawa Kazuhiko, ANNALS OF ONCOLOGY, 21, 35, 35,   2010 11
  • タモキシフェンによるMMPs発現及び浸潤阻害を介した転移抑制効果(Tamoxifen inhibits tumor metastasis through suppression of invasion and MMPs expression in mouse melanoma), 西田 升三, 金子 淳一, 椿 正寛, 松岡 寛, 木寺 康裕, 柳江 正嗣, 谷森 佳弘, 藤原 季美子, 森山 健三, 日本癌学会総会記事, 69回, 188, 189,   2010 08
  • DMFによるNF-kappaB抑制を介した転移抑制効果(Dimethylfumarate inhibit lung metastasis through the suppression of NF-kappaB signaling), 椿 正寛, 山添 譲, 松岡 寛, 金子 淳一, 木寺 康裕, 柳江 正嗣, 谷森 佳弘, 藤原 季美子, 森山 健三, 西田 升三, 日本癌学会総会記事, 69回, 420, 420,   2010 08
  • MIP-1 alphaはMEK/ERK経路を介して破骨細胞分化を誘導する(Macrophage inflammatory protein-1 alpha induces osteoclast formation by activation of MEK/ERK pathway), 西田 升三, 椿 正寛, 礒崎 美沙子, 礒野 藍, 佐藤 健太郎, 金子 淳一, 山添 譲, 木寺 康弘, 谷森 佳弘, 日本癌学会総会記事, 68回, 444, 444,   2009 08
  • タモキシフェンはPKC阻害を介して転移を抑制する(Tamoxifen inhibits tumor metastasis through the suppression of PKC in mouse melanoma), 椿 正寛, 松岡 寛, 礒崎 美沙子, 礒野 藍, 佐藤 健太郎, 金子 淳一, 荘子 夏緒里, 尾垣 光彦, 中村 春行, 柳江 正嗣, 西田 升三, 日本癌学会総会記事, 68回, 485, 485,   2009 08
  • StatinはRas/MEK/ERK経路を抑制することでがん細胞に対してアポトーシスを誘導する(Statin induces apoptosis in various tumor cells dependently on inhibition by Ras/MEK/ERK pathway), 椿 正寛, 西之坊 実里, 礒野 藍, 礒崎 美沙子, 佐藤 健太郎, 山添 譲, 谷森 佳弘, 木寺 康裕, 森山 健三, 西田 升三, 日本癌学会総会記事, 67回, 131, 131,   2008 09
  • MIP-1alphaによる骨髄間質細胞および骨芽細胞でのRANKL発現促進効果(MIP-1 alpha increases a RANKL expression in bone marrow stromal cells and osteoblasts through MAPK and PI3K/Akt pathway), 礒野 藍, 椿 正寛, 西之坊 実里, 礒崎 美沙子, 佐藤 健太郎, 山添 譲, 木寺 康裕, 谷森 佳弘, 森山 健三, 西田 升三, 日本癌学会総会記事, 67回, 149, 149,   2008 09
  • PKC阻害剤は悪性黒色腫における腫瘍浸潤を抑制する(PKC inhibitor suppresses tumor cell invasion in mouse melanoma cells), 礒崎 美沙子, 椿 正寛, 西之坊 実里, 礒野 藍, 佐藤 健太郎, 荘子 夏緒里, 尾垣 光彦, 中村 春行, 松岡 寛, 西田 升三, 日本癌学会総会記事, 67回, 186, 186,   2008 09
  • 窒素含有ビスホスホネート、YM529/ONO5920による骨髄腫でのMIP-1alpha分泌抑制(Nitrogen-containing bisphosphonate, YM529/ONO-5920, inhibits MIP-1 alpha secretion in myeloma cells), 西之坊 実里, 椿 正寛, 礒野 藍, 礒崎 美沙子, 佐藤 健太郎, 山添 譲, 谷森 佳弘, 木寺 康裕, 森山 健三, 西田 升三, 日本癌学会総会記事, 67回, 308, 309,   2008 09
  • PKC阻害剤はタイプIおよびタイプIVコラーゲンとの接着を阻害することで転移を抑制する(Protein kinase C inhibitor inhibited metastasis through suppression of cell adhesion to type I and type IV collagen), 佐藤 健太郎, 椿 正寛, 西之坊 実里, 礒野 藍, 礒崎 美沙子, 荘子 夏緒里, 尾垣 光彦, 中村 春行, 松岡 寛, 西田 升三, 日本癌学会総会記事, 67回, 367, 367,   2008 09
  • マウス骨髄間質細胞及び骨芽細胞においてマクロファージ炎症性蛋白質1α(MIP-1α)は核因子κBリガンドの受容体活性化因子(RANKL)の発現を増強する(Macrophage inflammatory protein-1alpha (MIP-1alpha) enhances a receptor activator of nuclear factor kappaB ligand (RANKL) expression in mouse bone marrow stromal cells and osteoblast), 西之坊 実里, 椿 正寛, 尾垣 光彦, 山添 譲, 藤原 季美子, 松岡 寛, 西浦 早織, 西田 升三, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 80回・30回, 4T18, 3,   2007 11
  • タンパク質キナーゼC阻害剤H7は細胞接着阻害を介してマウス黒色腫の肺転移を抑制する(Protein kinase C inhibitor, H7, inhibits lung metastasis in mouse melanoma through suppression of cell adhesion), 椿 正寛, 尾垣 光彦, 西之坊 実里, 藤原 季美子, 山添 譲, 森山 健三, 西田 升三, 日本癌学会総会記事, 66回, 147, 147,   2007 08
  • PKC阻害剤H7はERK1/2活性化の抑制を介して腫瘍浸潤を抑制する(The PKC inhibitor, H7, inhibits tumor cell invasion and through the suppression of ERK1/2 activation), 尾垣 光彦, 椿 正寛, 西之坊 実里, 藤原 季美子, 山添 譲, 森山 健三, 西田 升三, 日本癌学会総会記事, 66回, 147, 148,   2007 08
  • HL60細胞においてメバスタチンはリン酸化ERK1/2減少依存性にアポトーシスを誘導する(Mevastatin induces apoptosis in HL60 cells dependently on decrease in phosphorylated ERK1/2), 西之坊 実里, 椿 正寛, 尾垣 光彦, 藤原 季美子, 山添 譲, 森山 健三, 西田 升三, 日本癌学会総会記事, 66回, 366, 366,   2007 08
  • 新規ビスホスホネートYM529はリン酸化ERK1/2減少依存性にHL60細胞のアポトーシスを誘導する(A novel bisphosphonate, YM529, induces apoptosis in HL60 cells dependent on decrease in phosphorylated ERK1/2), 藤原 季美子, 椿 正寛, 尾垣 光彦, 西之坊 実里, 山添 譲, 森山 健三, 西田 升三, 日本癌学会総会記事, 66回, 366, 366,   2007 08
  • Apoptosis-inducing effect of mevastatin on HL60 cells and its mechanism, M Tsubaki, M Yanae, C Kato, Y Tanimori, Y Fujii, S Nishida, JOURNAL OF PHARMACOLOGICAL SCIENCES, 100, 126P, 126P,   2006
  • Nitrogen-containg bisphosphonate, YM529 inhibits RANKL expression in bone marrow stromal cell line ST2, C Kato, M Tsubaki, M Hoshino, A Namimatsu, H Uji, M Yanae, S Nishida, JOURNAL OF PHARMACOLOGICAL SCIENCES, 100, 71P, 71P,   2006
  • Nitrogen-containing bisphosphonate, YM529/ONO-5920 (a novel minodronic acid), inhibits RANKL expression in a cultured bone marrow stromal cell line ST2, S Nishida, M Tsubaki, M Hoshino, A Namimatsu, H Uji, S Yoshioka, Y Tanimori, M Yanae, M Iwaki, K Irimajiri, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 328, 1, 91, 97,   2005 03 , 10.1016/j.bbrc.2004.13.145
    Summary:Increase in bone resorption by osteoclasts can cause metabolic bone diseases, such as osteoporosis. Recent attention has been paid to the receptor activator of the NF-kappaB ligand (RANKL), an accelerator of osteoclast differentiation. RANKL is expressed on the bone marrow-derived stromal cell membrane and induces the differentiation of osteoclasts by binding to RANK expressed on the osteoclast precursor cell membrane. Since the inhibition of RANKL expression can lead to the inhibition of osteoclastic bone resorption, the clinical application of RANKL inhibition could be expected to have a major effect on metabolic bone disease therapy. In this study, we investigated whether or not YM529/ONO-5920, a nitrogen-containing bisphosphonate (a novel minodronic acid), inhibits RANKL expression in a bone marrow-derived stromal cell line (ST2 cells). Reverse transcription-polymerase chain reaction revealed that the administration of YM529/ONO-5920 to ST2 cells inhibited RANKL mRNA expression and reduced RANKL proteins as assessed by Western blot analysis. The inhibition of RANKL mRNA expression was reversed when geranylgeranyl pyrophosphate (GGPP), an intermediate in the mevalonate pathway, was used in combination. Furthermore, YM529/ ONO-5920 reduced phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2), and similarly, U0126, a mitogen-activated protein kinase kinase 1/2 inhibitor, inhibited RANKL expression. Pretreatment with GGPP reversed the YM529/ONO-5920-induced decrease in phosphorylation of ERK. Furthermore, YM529/ONO-5920 decreased TRAP-positive cells in co-culture of ST2 cells and an osteoclast cell line, C7 cells, and this decrease was inhibited by pretreatment with GGPP. This indicates that YM529/ONO-5920 inhibits GGPP biosynthesis in the mevalonate pathway and then signal transduction in the Ras-mitogen-activated protein kinase pathway, thereby inhibiting RANKL expression on ST2 cells. These results suggest a newly elucidated action of bisphosphonates in the inhibition of bone resorption. (C) 2005 Elsevier Inc. All rights reserved.
  • Mevastatin induces apoptosis in HL60 cells dependently on decrease in phosphorylated ERK, S Nishida, H Matsuoka, M Tsubaki, Y Tanimori, M Yanae, Y Fujii, M Iwaki, MOLECULAR AND CELLULAR BIOCHEMISTRY, 269, 1-2, 109, 114,   2005 01 , 10.1007/s11010-005-3086-0
    Summary:Mevastatin which is an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme in cholesterol synthesis, suppress cell proliferation and induce apoptosis. However, the molecular mechanism of apoptosis induction is not well understood. So, in the present study, we attempted to clarify the mechanism by which mevastatin induces apoptosis in HL60 cells. It was found that mevastatin induced apoptosis. At that time, we observed an increase in caspase-3 activity and morphological fragmentation of the nuclei. The apoptosis induced by mevastatin was not inhibited by the addition of farnesyl pyrophosphate (FPP), squalene, ubiquinone, and isopentenyladenine, but was inhibited by the addition of geranylgeranyl pyrophosphate (GGPP). When we examined the survival signals at the time of apoptotic induction, we also observed that the administration of mevastatin had caused a remarkable decrease in the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). However, other survival signals, such as nuclear factor kappa B (NF-kappaB), protein kinase B (Akt), and p38 mitogen-activated protein kinase (p38), exhibited no change. In addition, no quantitative change was observed in Bcl-2, which was an anti-apoptosis protein. It was also observed that apoptosis was induced when U0126, an MEK inhibitor, was added to the cells to inhibit ERK. These results suggested that mevastatin induced apoptosis when it inhibited GGPP biosynthesis and consequently decreased the level of phosphorylated ERK, which was a survival signal; moreover, at that time, there was no influence on NF-kappaB, Akt, p38, and Bcl-2. The results of this study also suggested that mevastatin could be used as an anticancer agent.
  • Diethyldithiocarbamate can induce two different type of death: Apoptosis and necrosis mediating the differential MAP kinase activation and redox regulation in HL60 cells, S Kimoto-Kinoshita, S Nishida, TT Tomura, MOLECULAR AND CELLULAR BIOCHEMISTRY, 265, 1-2, 123, 132,   2004 10 , 10.1023/B:MCBI.0000044366.32073.2d
    Summary:Although apoptosis and necrosis have been considered different pathways to cell death, only one compound induces both types of cell death. Diethyldithiocarbamate (DDC) has been shown to have antioxidant or prooxidant effects in several different systems. We observed in our present study that DDC induced not only apoptosis but also necrosis depending on its dosage in HL60 premyelocytic leukemia cells. Moreover, in hypoxia cell culture conditions, DDC-induced necrotic cells decreased but DDC-induced apoptosis continued. We investigated the DDC-induced different cell death mechanisms as they are correlated with reactive oxygen species (ROS). High-dose DDC-induced necrotic cell death is thought to depend on the increase of intracellular ROS, while low-dose DDC-induced apoptosis is thought to depend on changes of the intracellular redox state by the transporting of external metal ions. There was no sequential or quantitative change of Bcl-2 family proteins in DDC-induced apoptotic or necrotic pathways. However, the mitochondrial transmembrane potential was remarkably decreased in the DDC-induced necrosis. Finally, duration of c-Jun N-terminal kinase (JNK) activation resulted in different types of cell death.
  • Pretreatment with PKC inhibitor triggers TNF-alpha induced apoptosis in TNF-alpha-resistant B16 melanoma BL6 cells, S Nishida, S Yoshioka, S Kinoshita-Kimoto, M Kotani, M Tsubaki, Y Fujii, TT Tomura, K Irimajiri, LIFE SCIENCES, 74, 6, 781, 792,   2003 12 , 10.1016/j.lfs.2003.07.012
    Summary:Tumor necrosis factor alpha (TNF-alpha) modulates various events through several different pathways. Many tumor cells are resistant to this cytokine. Pretreatment of these cells with actinomycin D enhances TNF-alpha-induced apoptosis. In the present study, we investigated the mechanism of this enhancement and whether or not the apoptosis of TNF-alpha-resistant cancer cells can be induced by the inhibition of Protein kinase C (PKC). When TNF-alpha was added after inhibition of PKC by H7, apoptosis was observed, and companied with the activation of nuclear factor kappa B (NF-kappaB). After the inhibition of protein kinase B (Akt) by LY294002 or p38 mitogen-activated protein kinase (p38MAPK) by SB203580, the addition of TNF-alpha did not cause apoptosis. However, after the inhibition of MAPK/extracellular signal-regulated kinase kinase 1/2 (MEK1/2) with U0126, apoptosis was observed when TNF-alpha was added. In the Western blotting analysis, phosphorylation of MEK1/2 occurred at 60 minutes after the addition of TNF-alpha. However, it was noted that after pretreatment with H7, a significant decrease in phosphorylated MEKI/2 was observed. The present findings suggest that MEK1/2 plays an important role in TNF-alpha-resistance in TNF-alpha-resistant B16 melanoma BL6 cells. Furthermore, it was found that MEKI/2 is more important than NF-kappaB, Akt, and p38MAPK in anti-apoptotic PKC signaling and that TNF-alpha-resistance can be overcome by inhibiting MEKI/2. These results suggest the possibility of development of a new anticancer drug treatment. (C) 2003 Elsevier Inc. All rights reserved.
  • A new bisphosphonate, YM529 induces apoptosis in HL60 cells by decreasing phosphorylation of single survival signal ERK, S Nishida, Y Fujii, S Yoshioka, S Kikuichi, M Tsubaki, K Irimajiri, LIFE SCIENCES, 73, 21, 2655, 2664,   2003 10 , 10.1016/S0024-3205(03)00664-7
    Summary:It is believed that bisphosphonates (BPs) induce apoptosis in cells such as myeloma cells, as they inhibit prenylation of G-proteins. However, the details of the apoptosis-inducing mechanism remain obscure. In the present study, we attempted to clarify the mechanism by which YM529, a new bisphosphonate, induces apoptosis. YM529 induced cell deaths in HL60 cells in a concentration-dependent manner. At that time, we observed an increase in Caspase-3 activity and morphological fragmentation of the nuclei. We could confirm that these cell deaths were evidence of apoptosis. The apoptosis induced by YM529 was not inhibited by the addition of farnesyl pyrophosphate (FPP), but was by the addition of geranylgeranyl pyrophosphate (GGPP). When we examined the survival signals at the time of apoptotic induction, we also observed that the administration of YM529 caused a remarkable decrease in the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). However, other survival signals such as nuclear factor kappa B (NF-kappaB), protein kinase B (Akt), and p38 mitogen-activated protein kinase (p38) exhibited no change. In addition, no quantitative change was observed in Bcl-2, which is an anti-apoptosis protein. It was also observed that apoptosis was induced when U0126, an MEK inhibitor, was added to the cells to inhibit ERK. These results suggest that YM529, the new bisphosphonate, induced apoptosis when inhibit GGPP synthase and consequently decreased the levels of phosphorylated ERK, which is a survival signal; moreover, during this process, there is no influence on NF-kappaB, Akt, p38, and Bcl-2. The results of this study also suggest that YM529 can be used as an anticancer agent, in addition to its use as a therapeutic agent to treat osteoporosis. (C) 2003 Elsevier Inc. All rights reserved.
  • A new bisphosphonate, YM529 induces apoptosis in HL60 cells by decreasing phosphorylation of single survival signal ERK, S Nishida, Y Fujii, S Yoshioka, S Kikuichi, M Tsubaki, K Irimajiri, LIFE SCIENCES, 73, 21, 2655, 2664,   2003 10 , 10.1016/S0024-3205(03)00664-7
    Summary:It is believed that bisphosphonates (BPs) induce apoptosis in cells such as myeloma cells, as they inhibit prenylation of G-proteins. However, the details of the apoptosis-inducing mechanism remain obscure. In the present study, we attempted to clarify the mechanism by which YM529, a new bisphosphonate, induces apoptosis. YM529 induced cell deaths in HL60 cells in a concentration-dependent manner. At that time, we observed an increase in Caspase-3 activity and morphological fragmentation of the nuclei. We could confirm that these cell deaths were evidence of apoptosis. The apoptosis induced by YM529 was not inhibited by the addition of farnesyl pyrophosphate (FPP), but was by the addition of geranylgeranyl pyrophosphate (GGPP). When we examined the survival signals at the time of apoptotic induction, we also observed that the administration of YM529 caused a remarkable decrease in the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). However, other survival signals such as nuclear factor kappa B (NF-kappaB), protein kinase B (Akt), and p38 mitogen-activated protein kinase (p38) exhibited no change. In addition, no quantitative change was observed in Bcl-2, which is an anti-apoptosis protein. It was also observed that apoptosis was induced when U0126, an MEK inhibitor, was added to the cells to inhibit ERK. These results suggest that YM529, the new bisphosphonate, induced apoptosis when inhibit GGPP synthase and consequently decreased the levels of phosphorylated ERK, which is a survival signal; moreover, during this process, there is no influence on NF-kappaB, Akt, p38, and Bcl-2. The results of this study also suggest that YM529 can be used as an anticancer agent, in addition to its use as a therapeutic agent to treat osteoporosis. (C) 2003 Elsevier Inc. All rights reserved.
  • Induction of apoptosis in HL-60 cells treated with medicinal herbs, S Nishida, S Kikuichi, S Yoshioka, M Tsubaki, Y Fujii, H Matsuda, M Kubo, K Irimajiri, AMERICAN JOURNAL OF CHINESE MEDICINE, 31, 4, 551, 562,   2003 , 10.1142/S0192415X03001211
    Summary:In order to develop a new apoptosis inducer, we screened 22 crude drugs for their apoptosis-inducing activity. It was found that Glycyrrhiza uralensis, Cynomorium songaricum, Eucommia ulmoides, Phellodendron amurense, Cinnamomum cassia and Paeonia lactiflora induced the death of HL-60 cells. To investigate the mechanism of apoptosis induced by these six crude drugs, the mitochondrial transmembrane potential and the activity of caspase-3 were measured. Reduced mitochondrial transmembrane potentials within 12 hours after the administration of Glycyrrhiza uralensis, Cynomorium songaricum, Phellodendron amurense and Paeonia lactiflora, and within 24 hours after the administration of Eucommia ulmoides and Cinnamomum cassia were observed. All of the six apoptosis-inducing crude drugs increased caspase-3 activity within 12-36 hours after administration. After further examining the apoptosis-inducing activity of berberine, palmatine, panelofuroline and glycyrrhizin, which were the ingredients obtained from Phellodendron amurense, Glycyrrhiza uralensis and Paeonia lactiflora, it was found that only berberine could induce apoptosis. From these results, it was concluded that the apoptosis induced by the six crude drugs (Glycyrrhiza uralensis, Cynomorium songaricum, Eucommia ulmoides, Phellodendron amurense, Cinnamomum cassia and Paeonia lactiflora) occurred via the mitochondrial route and that the apoptosis-conducting mechanism acted through a cascade involving caspase-3.
  • Apoptosis-inducing effect of a new bisphosphonate, YM529, on various hematopoietic tumor cell lines, S Nishida, S Kikuichi, H Haga, S Yoshioka, M Tsubaki, K Fujii, K Irimajiri, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 26, 1, 96, 100,   2003 01 , 10.1248/bpb.26.96
    Summary:In recent years, it has been reported that bisphosphonates inhibited the cell cycle of myeloma cells to inhibit cell proliferation directly, and it was also reported that bisphosphonates induced apoptosis of myeloma cells in vitro. Recently, YM529 was developed as a new third-generation bisphosphonate. In our experiment, we investigated whether YM529 showed an antitumor effect on hematopoietic tumor cell lines other than myeloma, and we compared YM529 with YM175, which had a relatively more potent antitumor effect than that of existing bisphosphonates. We found that YM529 inhibited cell proliferation in various hematopoietic tumor cell lines (acute promyelocytic leukemia cell line HL-60, chronic myeloid leukemia cell line K562, histiocytic lymphoma cell line U937, lymphoblastic leukemia T cell line Jurkat, acute lymphoblastic leukemia T cell line MOLT-4, lymphoblastic leukemia B cell line CCRF-SB) including myeloma (myeloma cell line HS-Sultan) dose-dependently and time-dependently to a degree equivalent or superior to that in myeloma, and induced apoptosis at a lower concentration as compared with YM175. We confirmed many dead cells as well as apoptosis based on the detection of the nuclei with separate globular structure, the activation of caspase-3, and the decrease in mitochondrial transmembrane potential. Therefore, it is concluded that further utilization of YM529 can be expected against hematopoietic tumor cells in the future.
  • Apoptosis-inducing effect of a new bisphosphonate, YM529, on various hematopoietic tumor cell lines, S Nishida, S Kikuichi, H Haga, S Yoshioka, M Tsubaki, K Fujii, K Irimajiri, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 26, 1, 96, 100,   2003 01 , 10.1248/bpb.26.96
    Summary:In recent years, it has been reported that bisphosphonates inhibited the cell cycle of myeloma cells to inhibit cell proliferation directly, and it was also reported that bisphosphonates induced apoptosis of myeloma cells in vitro. Recently, YM529 was developed as a new third-generation bisphosphonate. In our experiment, we investigated whether YM529 showed an antitumor effect on hematopoietic tumor cell lines other than myeloma, and we compared YM529 with YM175, which had a relatively more potent antitumor effect than that of existing bisphosphonates. We found that YM529 inhibited cell proliferation in various hematopoietic tumor cell lines (acute promyelocytic leukemia cell line HL-60, chronic myeloid leukemia cell line K562, histiocytic lymphoma cell line U937, lymphoblastic leukemia T cell line Jurkat, acute lymphoblastic leukemia T cell line MOLT-4, lymphoblastic leukemia B cell line CCRF-SB) including myeloma (myeloma cell line HS-Sultan) dose-dependently and time-dependently to a degree equivalent or superior to that in myeloma, and induced apoptosis at a lower concentration as compared with YM175. We confirmed many dead cells as well as apoptosis based on the detection of the nuclei with separate globular structure, the activation of caspase-3, and the decrease in mitochondrial transmembrane potential. Therefore, it is concluded that further utilization of YM529 can be expected against hematopoietic tumor cells in the future.
  • Induction of apoptosis in HL-60 cells treated with medicinal herbs, S Nishida, S Kikuichi, S Yoshioka, M Tsubaki, Y Fujii, H Matsuda, M Kubo, K Irimajiri, AMERICAN JOURNAL OF CHINESE MEDICINE, 31, 4, 551, 562,   2003 , 10.1142/S0192415X03001211
    Summary:In order to develop a new apoptosis inducer, we screened 22 crude drugs for their apoptosis-inducing activity. It was found that Glycyrrhiza uralensis, Cynomorium songaricum, Eucommia ulmoides, Phellodendron amurense, Cinnamomum cassia and Paeonia lactiflora induced the death of HL-60 cells. To investigate the mechanism of apoptosis induced by these six crude drugs, the mitochondrial transmembrane potential and the activity of caspase-3 were measured. Reduced mitochondrial transmembrane potentials within 12 hours after the administration of Glycyrrhiza uralensis, Cynomorium songaricum, Phellodendron amurense and Paeonia lactiflora, and within 24 hours after the administration of Eucommia ulmoides and Cinnamomum cassia were observed. All of the six apoptosis-inducing crude drugs increased caspase-3 activity within 12-36 hours after administration. After further examining the apoptosis-inducing activity of berberine, palmatine, panelofuroline and glycyrrhizin, which were the ingredients obtained from Phellodendron amurense, Glycyrrhiza uralensis and Paeonia lactiflora, it was found that only berberine could induce apoptosis. From these results, it was concluded that the apoptosis induced by the six crude drugs (Glycyrrhiza uralensis, Cynomorium songaricum, Eucommia ulmoides, Phellodendron amurense, Cinnamomum cassia and Paeonia lactiflora) occurred via the mitochondrial route and that the apoptosis-conducting mechanism acted through a cascade involving caspase-3.
  • Histopathological findings of human thyroid tumors and dynamic MRI, T Kusunoki, K Murata, S Nishida, T Tomura, M Inoue, AURIS NASUS LARYNX, 29, 4, 357, 360,   2002 10 , 10.1016/S0385-8146(02)00029-9
    Summary:Objective: Time intensity curves for gadolinium-diethylenetriaminepentaceticacid (Gd-DTPA) enhanced magnetic resonance imaging (MRI), namely dynamic MRI, were determined for thyroid diseases, and compared with findings of histopathologic examination. Methods: Time intensity curves for the thyroid tumors were determined. Three different patterns of time intensity curves were observed: a rapid washout pattern, a delayed washout pattern and no change. Cell proliferating activities of thyroid tumors were estimated immunohistochemically by proliferating cell nuclear antigen (PCNA). Results: In most of malignant diseases and a few benign diseases that had marked cell proliferative activity with showing more than 20% on labeling indexes of PCNA, the time intensity curve displayed the delayed washout pattern, in which intensity was above half-maximal value within 10 min after injection Gd-DTPA. Almost all benign diseases and a few well differentiated carcinomas displayed the rapid washout pattern, in which intensity was decreased to lower than half of peak grade within 10 min following injection and showed from 10 to 20% on labeling indexes of PCNA. Benign diseases that displayed no change of time intensity curve did not show PCNA positive cells. Conclusion: These findings suggested that the time intensity curve obtained from dynamic MRI might indicate cell proliferating activity of thyroid tumors. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
  • Change of Cu,Zn-superoxide dismutase activity of guinea pig lung in experimental asthma, S Nishida, K Teramoto, S Kimoto-Kinoshita, Y Tohda, S Nakajima, TT Tomura, K Irimajiri, FREE RADICAL RESEARCH, 36, 6, 601, 606,   2002 06 , 10.1080/10715760290029074
    Summary:Correlation between the level of reactive oxygen species (ROS) generated by airway inflammatory cells and superoxide dismutase (SOD) activity of pulmonary tissue during an asthma attach was investigated in a guinea pig model of allergic asthma. In addition, the influence of SOD inhibition by diethyldithiocarbamate (DDC, Cu-chelating agent) on the airway was investigated in terms of pulmonary function during an asthma attach. Relative to controls, the capacity of bronchoalveolar lavage fluid (BAL) cells to release ROS was significantly increased in guinea pigs sensitized with ovalbumin (OA) as the antigen, and significantly increased in guinea pigs with an asthma attack provoked by the inhalation of OA. SOD activity was increased significantly in the antigen-sensitized group. The asthma provocation group showed a tendency for increase in total SOD activity, compared with the sensitization group, whose increase was dependent on the increase in copper, zinc-SOD (Cu, Zn-SOD) activity. Pretreatment with DDC increased the severity and duration of the asthma attack. These results were indicated that Cu, Zn-SOD was closely involved in the asthma process, particularly in the scavenging of oxygen radicals secreted from BAL cells.
  • Change of Cu,Zn-superoxide dismutase activity of guinea pig lung in experimental asthma, S Nishida, K Teramoto, S Kimoto-Kinoshita, Y Tohda, S Nakajima, TT Tomura, K Irimajiri, FREE RADICAL RESEARCH, 36, 6, 601, 606,   2002 06 , 10.1080/10715760290029074
    Summary:Correlation between the level of reactive oxygen species (ROS) generated by airway inflammatory cells and superoxide dismutase (SOD) activity of pulmonary tissue during an asthma attach was investigated in a guinea pig model of allergic asthma. In addition, the influence of SOD inhibition by diethyldithiocarbamate (DDC, Cu-chelating agent) on the airway was investigated in terms of pulmonary function during an asthma attach. Relative to controls, the capacity of bronchoalveolar lavage fluid (BAL) cells to release ROS was significantly increased in guinea pigs sensitized with ovalbumin (OA) as the antigen, and significantly increased in guinea pigs with an asthma attack provoked by the inhalation of OA. SOD activity was increased significantly in the antigen-sensitized group. The asthma provocation group showed a tendency for increase in total SOD activity, compared with the sensitization group, whose increase was dependent on the increase in copper, zinc-SOD (Cu, Zn-SOD) activity. Pretreatment with DDC increased the severity and duration of the asthma attack. These results were indicated that Cu, Zn-SOD was closely involved in the asthma process, particularly in the scavenging of oxygen radicals secreted from BAL cells.
  • Correlation between morphology of colonies formed by human thyroid tumor cells in collagen gel culture and invasive ability, T Kusunoki, S Nishida, K Murata, T Tomura, THYROID, 12, 4, 281, 286,   2002 04
    Summary:Two different types of colonies from human thyroid papillary carcinomas were observed in Collagen gel culture. One was a branching type, with many three-dimensional outgrowths. The other was a spherical type, with no outgrowths. As viewed by electron microscopy, the tumor cells of spherical-type colonies had more microvilli on their surfaces than did cells in the branching-type colonies. Histochemically, the tumor cells of spherical-type colonies had stronger positive reactions than did tumor cells of branching-type colonies. This suggests that tumor cells of spherical-type colonies have a higher potential for dissemination and invasion than do tumor cells of branching-type colonies. In fact, cases with higher percentages of spherical-type colonies could show extracapsular invasion histopathologically. Our results indicate that collagen gel culture is a useful method for studying the heterogeneity of tumor cells as well as their invasive ability in vitro.
  • Protease-activated receptor-2 (PAR-2) in the rat gastric mucosa: immunolocalization and facilitation of pepsin/pepsinogen secretion, N Kawao, Y Sakaguchi, A Tagome, R Kuroda, S Nishida, K Irimajiri, H Nishikawa, K Kawai, MD Hollenberg, A Kawabata, BRITISH JOURNAL OF PHARMACOLOGY, 135, 5, 1292, 1296,   2002 03 , 10.1038/sj.bjp.0704562
    Summary:1 Agonists of protease-activated receptor-2 (PAR-2) trigger neurally mediated mucus secretion accompanied by mucosal cytoprotection in the stomach. The present study immunolocalized PAR-2 in the rat gastric mucosa and examined if PAR-2 could modulate pepsin/pepsinogen secretion in rats. 2 PAR-2-like immunoreactivity was abundant in the deep regions of gastric mucosa, especially in chief cells. 3 The PAR-2 agonist SLIGRL-NH2, but not the control peptide LSIGRL-NH2 adininistered i.v. repeatedly at 0.3-1 mumol kg(-1), four times in total, significantly facilitated gastric pepsin secretion, although a single dose produced no significant effect. 4 The PAR-2-mediated gastric pepsin secretion was resistant to omeprazole, N-G-nitro-L-arginine methyl ester (L-NAME) or atropine, and also to ablation of sensory neurons by capsaicin. 5 Our study thus provides novel evidence that PAR-2 is localized in mucosal chief cells and facilitates gastric pepsin secretion in the rats. most probably by a direct mechanism. British Journal of Pharmacology (2002).
  • Type IV collagen, type IV collagenase activity and ability of cell proliferation in human thyroid tumours, Takeshi Kusunoki, Shozo Nishida, Saori Kimoto-Kinoshita, Kiyotaka Murata, Takao Satou, Takanori Tomura, Asian Journal of Surgery, 25, 4, 304, 308,   2002 , 10.1016/S1015-9584(09)60196-2
    Summary:BACKGROUND: The processes of malignant tumour invasion and metastasis are known to include the destruction of cell stroma and vascular basement membrane. It has been suggested that type IV collagenase degrades type IV collagen, a main component of the basement membrane. METHODS: In our study, type IV collagenase activity in human thyroid tumours was measured by the Liotta method. The degree of destruction of diseased regions of thyroid tumours was immunohistochemically determined by anti-type IV collagen antibody staining. Cell proliferation in the tumours was estimated using anti-proliferating cell nuclear antigen (PCNA) and epidermal growth factor receptor (EGFR). RESULTS: T4 thyroid carcinomas with higher type IV collagenase activity and very weak type IV discontinuous immunostaining for type IV collagen of follicular basement membranes, exhibited many PCNA or EGFR positive cells. In benign tumours, normofollicular- or macrofollicular-type tumours with low type IV collagenase activity showed few PCNA and EGFR positive cells and intact type IV collagen of basement membranes, as seen in normal thyroids. Conversely, an atypical adenoma with higher type IV collagenase activity showed many PCNA and EGFR positive cells and weak type IV discontinuous immunostaining for type IV collagen, as in thyroid carcinomas. CONCLUSION: These findings suggest that staining for type IV collagen and type IV collagenase activity reflect the ability of cell proliferation, and help predict the aggressiveness of invasion and metastasis in human thyroid tumours.
  • Correlation between morphology of colonies formed by human thyroid tumor cells in collagen gel culture and invasive ability, Takeshi Kusunoki, Shozo Nishida, Kiyotaka Murata, Takanori Tomura, Thyroid, 12, 4, 281, 286,   2002 , 10.1089/10507250252949397
    Summary:Two different types of colonies from human thyroid papillary carcinomas were observed in collagen gel culture. One was a branching type, with many three-dimensional outgrowths. The other was a spherical type, with no outgrowths. As viewed by electron microscopy, the tumor cells of spherical-type colonies had more microvilli on their surfaces than did cells in the branching-type colonies. Histochemically, the tumor cells of spherical-type colonies had stronger positive reactions than did tumor cells of branching-type colonies. This suggests that tumor cells of spherical-type colonies have a higher potential for dissemination and invasion than do tumor cells of branching-type colonies. In fact, cases with higher percentages of spherical-type colonies could show extracapsular invasion histopathologically. Our results indicate that collagen gel culture is a useful method for studying the heterogeneity of tumor cells as well as their invasive ability in vitro.
  • Morphological and functional differentiation of human thyroid cells in collagen gel culture, T Kusunoki, S Nishida, M Koezuka, K Murata, T Tomura, AURIS NASUS LARYNX, 28, 4, 333, 338,   2001 11 , 10.1016/S0385-8146(01)00109-2
    Summary:Objective: cultured human thyroid cells in collagen gel culture were examined on cell morphology and the production of thyroglobulin (Tg), triiodothyronine (T3) and thyroxine (T4) which are components of functional differentiation. Methods: thyroid cells obtained from normal human thyroid tissues (four cases), follicular adenoma tissues (three cases). papillary carcinoma tissues (three cases), and follicular carcinoma tissue (one case), were cultured in collagen gel. Then these cultured cells were observed on cellular morphology and production of Tg, T3 and T4. Moreover, changes in morphological characteristics and production of Tg, T3 and T4 induced by addition of thyroid stimulating hormone (TSH) and epidermal growth factor (EGF) to medium in collagen gel culture were determined. Results: normal and tumor cells in collagen gel culture formed colonies and follicles with Tg production, similar to in vivo-like three-dimensional cellular structures and functions. Normal thyroid cells stimulated TSH induced more Tg and produced morphological changes, i.e. enlarged follicular lumens and increased the height of follicular cells, but did not promote cell proliferation. Reversely, normal thyroid cells stimulated with EGF promoted cell proliferation, but did not change morphological findings and did not increase production of Tg, T3 and T4, Conclusion: these findings suggest that collagen gel culture is useful for observing the effects of stimulation by cell growth factor on the morphological and functional differentiation of human thyroid cells. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.
  • Morphological and functional differentiation of human thyroid cells in collagen gel culture, T Kusunoki, S Nishida, M Koezuka, K Murata, T Tomura, AURIS NASUS LARYNX, 28, 4, 333, 338,   2001 11 , 10.1016/S0385-8146(01)00109-2
    Summary:Objective: cultured human thyroid cells in collagen gel culture were examined on cell morphology and the production of thyroglobulin (Tg), triiodothyronine (T3) and thyroxine (T4) which are components of functional differentiation. Methods: thyroid cells obtained from normal human thyroid tissues (four cases), follicular adenoma tissues (three cases). papillary carcinoma tissues (three cases), and follicular carcinoma tissue (one case), were cultured in collagen gel. Then these cultured cells were observed on cellular morphology and production of Tg, T3 and T4. Moreover, changes in morphological characteristics and production of Tg, T3 and T4 induced by addition of thyroid stimulating hormone (TSH) and epidermal growth factor (EGF) to medium in collagen gel culture were determined. Results: normal and tumor cells in collagen gel culture formed colonies and follicles with Tg production, similar to in vivo-like three-dimensional cellular structures and functions. Normal thyroid cells stimulated TSH induced more Tg and produced morphological changes, i.e. enlarged follicular lumens and increased the height of follicular cells, but did not promote cell proliferation. Reversely, normal thyroid cells stimulated with EGF promoted cell proliferation, but did not change morphological findings and did not increase production of Tg, T3 and T4, Conclusion: these findings suggest that collagen gel culture is useful for observing the effects of stimulation by cell growth factor on the morphological and functional differentiation of human thyroid cells. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.
  • Cathepsin L activity and its inhibitor in human otitis media, T Kusunoki, S Nishida, K Murata, K Kobashi, H Nakatani, T Hiwasa, T Tomura, JOURNAL OF OTOLARYNGOLOGY, 30, 3, 157, 161,   2001 06
    Summary:Objective: We examined cathepsin L activity, expression of cystatin A, and copper- and zinc-containing superoxide dismutase in human chronic otitis media. The relationships of our findings to clinical findings (e.g., grade of bone destruction) were also studied. Design: Retrospective basic and clinical study. Setting: Department of Otolaryngology and First Department of Biochemistry, Kinki University School of Medicine, Osaka, Japan. Method: The human middle ear tissues evaluated in this study were surgically obtained from seven patients with cholesteatoma epithelium, three patients with granulation tissues in cholesteatoma, three patients with granulation tissues in noncholesteatoma, and three patients with intact mucous membrane of the middle ear. Main Outcome Measures: Cathepsin L activities in cholesteatoma epithelium, granulation tissues in cholesteatoma, or granulation tissues in noncholesteatoma were measured using Barrett's method. Cystatin A expressions were observed by Western blot analysis. Copper- and zinc-containing superoxide dismutase in cholesteatoma was examined immunohistochemically. Results: Mean cathepsin L activity was higher in diseased tissues than in intact mucous membranes of the middle ear. Granulation tissues with high cathepsin L activity resulted in extensive bone destruction in both cholesteatomas and non-cholesteatomas of the middle ear. All cases with intact mucous membrane of the middle ear exhibited no expression of cystatin A. Seven of 10 cases with diseased tissues expressed cystatin A in cholesteatoma epithelium, granulation tissues in cholesteatoma, or granulation tissues in noncholesteatoma. No relationships were found between cystatin A expression and grade of cathepsin L activity. Copper- and zinc-containing superoxide dismutase was more strongly positive in cholesteatoma epithelium regions than in granulation tissues. Conclusion: These results suggest that copper- and zinc-containing superoxide dismutase in cholesteatoma epithelium prevents complications by suppressing cathepsin L activity.
  • Cathepsin L activity and its inhibitor in human otitis media, T Kusunoki, S Nishida, K Murata, K Kobashi, H Nakatani, T Hiwasa, T Tomura, JOURNAL OF OTOLARYNGOLOGY, 30, 3, 157, 161,   2001 06
    Summary:Objective: We examined cathepsin L activity, expression of cystatin A, and copper- and zinc-containing superoxide dismutase in human chronic otitis media. The relationships of our findings to clinical findings (e.g., grade of bone destruction) were also studied. Design: Retrospective basic and clinical study. Setting: Department of Otolaryngology and First Department of Biochemistry, Kinki University School of Medicine, Osaka, Japan. Method: The human middle ear tissues evaluated in this study were surgically obtained from seven patients with cholesteatoma epithelium, three patients with granulation tissues in cholesteatoma, three patients with granulation tissues in noncholesteatoma, and three patients with intact mucous membrane of the middle ear. Main Outcome Measures: Cathepsin L activities in cholesteatoma epithelium, granulation tissues in cholesteatoma, or granulation tissues in noncholesteatoma were measured using Barrett's method. Cystatin A expressions were observed by Western blot analysis. Copper- and zinc-containing superoxide dismutase in cholesteatoma was examined immunohistochemically. Results: Mean cathepsin L activity was higher in diseased tissues than in intact mucous membranes of the middle ear. Granulation tissues with high cathepsin L activity resulted in extensive bone destruction in both cholesteatomas and non-cholesteatomas of the middle ear. All cases with intact mucous membrane of the middle ear exhibited no expression of cystatin A. Seven of 10 cases with diseased tissues expressed cystatin A in cholesteatoma epithelium, granulation tissues in cholesteatoma, or granulation tissues in noncholesteatoma. No relationships were found between cystatin A expression and grade of cathepsin L activity. Copper- and zinc-containing superoxide dismutase was more strongly positive in cholesteatoma epithelium regions than in granulation tissues. Conclusion: These results suggest that copper- and zinc-containing superoxide dismutase in cholesteatoma epithelium prevents complications by suppressing cathepsin L activity.
  • Decrease of endothelial nitric oxide synthase in stroke-prone spontaneously hypertensive rat cerebral cortex, S Kimoto-Kinoshita, S Nishida, TT Tomura, NEUROSCIENCE LETTERS, 288, 2, 103, 106,   2000 07 , 10.1016/S0304-3940(00)01227-1
    Summary:We reported in our previous study that constitutive nitric oxide synthase (cNOS) activity significantly decreased and oxygen stress increased in the 31-week-old stroke-prone spontaneously hypertensive rat (SHRSP) cerebral cortex (CC). In the present study we examined the protein amount of two cNOS isoforms, the neuronal and the endothelial types, in SHRSP CC using Western blot analysis. Although no significant difference was observed in the amount of neuronal NOS (nNOS) protein, endothelial NOS (eNOS) protein prominently decreased in 31-week-old SHRSP CC compared to age-matched Wistar Kyoto rat and 15-week-old SHRSP. In rats at this age, we also observed a large quantity of albumin in the protein amount. However, the protein amount of heat shock protein 70, which is a molecular chaperon and a marker of injury, showed no significant changes. These results indicate that the alteration of eNOS but not of nNOS protein would be more closely associated with the development of stroke in SHRSP. (C) 2000 Published by Elsevier Science Ireland Ltd.
  • Decrease of endothelial nitric oxide synthase in stroke-prone spontaneously hypertensive rat cerebral cortex, S Kimoto-Kinoshita, S Nishida, TT Tomura, NEUROSCIENCE LETTERS, 288, 2, 103, 106,   2000 07 , 10.1016/S0304-3940(00)01227-1
    Summary:We reported in our previous study that constitutive nitric oxide synthase (cNOS) activity significantly decreased and oxygen stress increased in the 31-week-old stroke-prone spontaneously hypertensive rat (SHRSP) cerebral cortex (CC). In the present study we examined the protein amount of two cNOS isoforms, the neuronal and the endothelial types, in SHRSP CC using Western blot analysis. Although no significant difference was observed in the amount of neuronal NOS (nNOS) protein, endothelial NOS (eNOS) protein prominently decreased in 31-week-old SHRSP CC compared to age-matched Wistar Kyoto rat and 15-week-old SHRSP. In rats at this age, we also observed a large quantity of albumin in the protein amount. However, the protein amount of heat shock protein 70, which is a molecular chaperon and a marker of injury, showed no significant changes. These results indicate that the alteration of eNOS but not of nNOS protein would be more closely associated with the development of stroke in SHRSP. (C) 2000 Published by Elsevier Science Ireland Ltd.
  • Age-related change of antioxidant capacities in the cerebral cortex and hippocampus of stroke-prone spontaneously hypertensive rats, S Kimoto-Kinoshita, S Nishida, TT Tomura, NEUROSCIENCE LETTERS, 273, 1, 41, 44,   1999 09 , 10.1016/S0304-3940(99)00623-0
    Summary:Regional distribution and age-related change of Mn-, Cu/Zn-superoxide dismutase (SOD) and constitutive type of nitric oxide synthase (NOS) activities in the brain were determined using stroke-prone spontaneously hypertensive rats (SHRSP). In the hippocampus (HIP), Mn- and Cu/Zn-SOD activities in SHRSP of 31-week-old were significantly lower than those of 15-week-old or normotensive rats (WKY). From Mn-SOD immunohistochemical staining of several subfields of the HIP, our results suggested that SHRSP hippocampal CA1 was more vulnerable to oxidative stress compared with WKY and other subfields. In the 31-week-old SHRSP cerebral cortex (CC), the activities of Mn-, Cu/Zn-SOD and NOS were significantly lower than those in WKY. At this age, most of the SHRSP developed cerebral injuries. These observations indicated that hypertensive vascular disease observed in the SHRSP CC resulted from the decreased antioxidant capacity that is closely associated with the development of stroke and, in turn, shortened life span. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.
  • Malignancies of human thyroid tumors and dynamic magnetic resonance imaging (MRI), T. Kusunoki, K. Murata, H. Hosoi, S. Nishida, T. Tomura, M. Inoue, Auris Nasus Larynx, 25, 4, 419, 424,   1998 12 , 10.1016/S0385-8146(98)00017-0
    Summary:Time intensity curves for gadolinium-diethylene triaminepentacetic acid (Gd-DTPA) enhanced magnetic resonance imaging (MRI), namely dynamic MRI, were determined for thyroid diseases and compared with findings of histopathologic examination. Time intensity curves for solid lesions were determined, excluding cases with secondary changes such as calcification, hemorrhage, necrosis and fibrosis. Three different patterns of time intensity curves were observed: rapid washout, delayed washout and no change. In our previous study, malignant grades of thyroid tumors were estimated immunohistochemically by epidermal growth factor receptor (EGFR) antibody. In most of malignant diseases and a few benign diseases that had marked cell proliferative activity with staining EGFR strongly, the time intensity curve displayed a delayed washout pattern, in which intensity was above 1/2- maximal value within 10 min after injection Gd-DTPA. Almost all benign diseases and a few well differentiated carcinomas displayed a rapid washout pastern, in which intensity was decreased to lower than 1/2 of peak grade within 10 min following injection and showed staining EGFR weakly. Benign diseases showing no change of time intensity curve, did not almost show aEGFR positive cell. These findings suggested that the time intensity curve obtained from dynamic MRI might indicate differentiated grades and cell proliferating activity of thyroid tumors.
  • Induction of manganese superoxide dismutase by thyroid stimulating hormone in rat thyroid cells, S Nishida, T Nakano, S Kimoto, T Kusunoki, K Suzuki, N Taniguchi, K Murata, TT Tomura, FEBS LETTERS, 416, 1, 69, 71,   1997 10 , 10.1016/S0014-5793(97)01171-X
    Summary:Alterations in the superoxide dismutase (SOD) content of thyroid tissues occurring in association with thyroid dysfunction have been reported, In this study, the Mn-SOD content was found to increase in thyroid tissues of rats administered thyroid stimulating hormone (TSH) and in thyrocytes cultured in medium supplemented with TSH, Furthermore, in the thyroid glands of rats whose serum TSH level was elevated by inhibiting the synthesis of T3 and T4 by 6-methyl-2-thiouracil, the Mn-SOD increased as the TSH concentration increased, In the cultured thyrocytes, the increase in Mn-SOD induced by TSH was inhibited by the C-kinase inhibitor H7. These findings suggest the induction of Mn-SOD by TSH in thyroid cells and point to a role of C-kinase in this process, thereby indicating that a close relationship exists between the serum TSH level and the change in Mn-SOD content in thyrocytes with thyroid dysfunction, (C) 1997 Federation of European Biochemical Societies.
  • Enhancement of TNF-alpha induced antitumor activity by inhibition of superoxide dismutase induction, Cell structure and function, 22, 800, 801,   1997
  • コラーゲン・ゲル培養でのヒト甲状腺細胞のサイログロブリン,甲状腺ホルモン産生能の検討, 楠 威志, 西田 升三, 肥塚 正博, Auris・Nasus・Larynx, 22, 2, 120, 127,   1995 07
    Summary:ヒト正常甲状腺組織21症例,ヒト甲状腺病変組織(バセドウ病1例,濾胞腺腫4例,乳頭癌11例,濾胞癌1例)より得られた甲状腺細胞をコラーゲン・ゲル培養を行い,Tg,T3,T4産生能を観察した.Tg陽性を示したものは24例中20例に認め,病理組織診別にみてもTg陽性率にはほとんど差がなかった.T3,T4陽性を示した症例はそれぞれ3,4例と分化癌のみにみられた.TSH添加群では正常甲状腺細胞のみに,Tg,T4産生を示すコロニーの出現率が上昇し,形態的には濾胞腔の拡大および濾胞上皮細胞の丈の増大がみとめられた.以上のように,コラーゲン・ゲル培養中の甲状腺細胞はより生体に近い所見を示している
  • ヒト甲状腺腫内のカテプシンB活性の検討, 楠 威志, 西田 升三, 中野 貴之, Auris・Nasus・Larynx, 22, 1, 43, 48,   1995 04
    Summary:切除した甲状腺組織を用いて,カテプシンB活性を測定し,母腫瘍の病理組織診断および転移,浸潤能さらには組織内1型および4型コラーゲン分解能についても対比検討した。カテプシンB活性の平均値は,良性および悪性病変群が正常甲状腺群よりも有意に高値を示した。そのなかでも,被膜外浸潤,リンパ節転移および強い細胞増殖を認める癌症例ではより高値を示した。カテプシンBで高値を認めた症例の組織内1型および4型コラーゲン分解能も高い傾向を示した
  • Productivity of Thyroglobulin and Thyroid Hormone on Human Thyroid Cell in Collagen Gel Culture, Takeshi Kusunoki, Shozo Nishida, Masahiro Koezuka, Kiyotaka Murata, Takanori Tomura, Auris Nasus Larynx, 22, 2, 120, 127,   1995 , 10.1016/S0385-8146(12)80110-6
    Summary:Thyroid cells obtained from 21 normal human thyroid tissue samples and 17 tissue samples from diseased thyroid, including one of Graves disease, 4 with follicular adenoma, 11 with papillary carcinoma, and 1 with follicular carcinoma, were cultured in collagen gel, and ability to produce thyroglobulin (Tg), triiodothyronine (T3) and thyroxine (T4) was determined. Changes in morphological characteristics and production of Tg, T3, and T4 induced by addition of thyroid stimulating hormone (TSH) to medium in collagen gel culture were also determined. Twenty of all cases exhibited positive reaction for Tg. No relationship was found between rate of positivity for Tg and pathologic diagnosis. Three with carcinoma showed positive reaction for T3, and 4 with carcinoma showed positive reaction for T4. Only for normal thyroid cells did addition of TSH to medium induce increase the percentage of colonies producing Tg or T4 and morphological changes including an enlarged follicular lumen and increase in the height of columnar epithelium. These findings suggest that thyroid cells in collagen culture develop in an in vivo-like fashion. In conclusion, collagen has important effects on cellular differentiation when included in extracellular matrix. © 1995, Society for Promotion of International Otorhinolaryngology (SPIO). All rights reserved.
  • REGIONAL DISTRIBUTION OF SUPEROXIDE DISMUTASE IN THE BRAIN AND MYOCARDIUM OF THE STROKE‐PRONE SPONTANEOUSLY HYPERTENSIVE RAT, S. Kimoto, S. Nishida, K. Funasaka, T. Nakano, K. Teramoto, T. T. Tomura, Clinical and Experimental Pharmacology and Physiology, 22, S160, S161,   1995 , 10.1111/j.1440-1681.1995.tb02863.x
    Summary:1. Enzyme activities and contents of manganese and copperzinc superoxide dismutase (Mn‐, Cu/Zn‐SOD) and oxygen free‐radical scavengers were determined in the myocardium (right, left ventricle) and brain (cerebral cortex, hippocampus) of 15 and 31 week old stroke‐prone spontaneously hypertensive rats (SHRSP). 2. In 15 week old SHRSP myocardium, both Mn‐ and Cu/Zn‐SOD activities were higher but in 31 week old SHRSP, these were lower than that in Wistar‐Kyoto (WKY) rats. Further, correlation between Mn‐SOD content and activity in 31 week old SHRSP myocardium showed that specific activity was lower than that in WKY. 3. In 15 and 31 week old SHRSP cerebral cortex and hippocampus, SOD content and activity showed a tendency to be lower than that in WKY. 4. These results indicate that enzymatically inactive or low‐active Mn‐SOD protein exists in SHRSP myocardium, and that the alteration of SOD may be one of the causative factors for the vulnerability of the myocardium and brain against O2‐radicals. Copyright © 1995, Wiley Blackwell. All rights reserved
  • シリコンチューブ内の切除ラット坐骨神経の再生に及ぼすコラーゲンゲル基質の影響に関する形態学的研究, 佐藤 隆夫, 西田 升三, 蛭間 真悟, Acta Pathologica Japonica, 36, 2, 199, 208,   1986 02
    Summary:坐骨神経を5 mm切除し,その近位と遠位端を5 mm離してシリコンチューブに入れ,その間にコラーゲンゲル基質を入れ,その神経再生に及ぼす影響を見た.間に何も入れない対照群に較べて,著しい神経の成長が認められたが,線維芽細胞,シュワン細胞を含む細胞は少なかった.また神経の成長も対照群に比べてコラーゲン基質の入ったチューブではより方向がそろっていた.シリコンチューブ内における末梢神経の再生がコラーゲン基質の使用により改善されることが示された
  • IMMUNOHISTOCHEMICAL CHANGES OF THE ISCHEMIC HEART-MUSCLE FIBERS WITH ANTIACTIN ANTIBODIES, S NISHIDA, M MAEDA, E KADOTA, S HASHIMOTO, ACTA HISTOCHEMICA ET CYTOCHEMICA, 18, 6, 660, 660,   1985
  • EXPERIMENTAL INVESTIGATION OF THE REACTIVITY OF LECTIN BINDING TO THE NORMAL VASCULAR ENDOTHELIUM, S NISHIDA, M MAEDA, E KADOTA, K TANJI, S HASHIMOTO, F AKAI, ACTA HISTOCHEMICA ET CYTOCHEMICA, 17, 6, 716, 716,   1984

Research Grants & Projects

  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Identification of imatinib resistance factor and the therapeutic strategies to overcome resistance in chronic myeloid leukemia, To investigate the underlying mechanisms associated with resistance to imatinib, we established imatinib-resistant chronic myeloid leukemia (CML) cell lines. The resistant cell lines activation of MET. In addition, MET inhibitor reversed the imatinib-resistance of the drug-resistant cell lines. These findings suggest that MET inhibitors are potentially useful as an imatinib-resistance CML agent for the treatment of CML cells. As well, These results are summarized as section of presented paper.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Chronic stress state promotes cancer metastatic ability, We investigated whether chronic stress influences on cancer metastatic ability using a lung cancer metastasis model B16BL6 cell. As a result, we obtained the following findings. 1) The cancer metastasis and the tumor growth in B16BL6 cell were promoted in chronically stressed mice. 2) The number of NK cell and alveolar macrophage and the phagocytic activity of the macrophage in the chronically stressed mice were decreased. It is suggested from these results suggest that the cancer metastatic ability of B16BL6 cell in chronically stressed mice is prompted so that the immunocompetence of the alveolar macrophage is decreased in the chronically stressed mice.
  • Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), SPINAL ISCHEMIA, ITS MECHANISM AND STRATEGY OF TREATMENT-A NEW MODEL AND GLIAL REACTION AND INDUCTION, 1) The light and electron microscopic localization of single strand DNA protein, a marker of apoptosis was studied. Immunoreaction was recognized in the nucleus without morphological changes 1 day after ischemic insult. Degradation of single strand DNA can occur during delayed neuronal death, preceding the appearance of double strand DNA breaks.(Neurosci Lett 240 : 69, 1998) 2) DDC induced apoptosis in a morphological nature in PC12 cells, and led to down -regulation of Cu/Zn-SOD activity, then reduced intracellular HィイD22ィエD2OィイD22ィエD2. A hypoxia prevented the cells from DDC-induced apoptosis. These data confirmed OィイD22ィエD2 induced apoptosis in PC12 cells. Nitro-oxide-synthetase, Fe-chelator did not prevent the apoptosis. Peroxinitrate and also OH did not contribute DDC-induced apoptosis. NGF and NAC protected the apoptosis. DDC did not activate NF-kB. We concluded that NAC reduced DDC-induced apoptosis by interception of an unknown direct signal pathway, by which OィイD22ィエD2 induced apoptosis.
  • Analysis of the mechanism of tumor metastasis and discovery of agents for tumor metastasis
  • Induction of apoptosis induced by Bisphosphonate in hematopoietic cells
  • Discovery of anti-osteolytic agents and agents for anti-osteolytic metastasis