KINDAI UNIVERSITY


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ISHIWATA Shunji

Profile

FacultyDepartment of Pharmacy / Graduate School of Medicine
PositionAssociate Professor
Degree
Commentator Guidehttps://www.kindai.ac.jp/meikan/767-ishiwata-shunji.html
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Last Updated :2020/08/10

Research Activities

Published Papers

  • Investigation of the levels of Ifosfamide vaporized from powder and solution, ISHIWATA Shunji, Japanese Journal of Occupational Medicine and Traumatology, Japanese Journal of Occupational Medicine and Traumatology, 67, 95 - 99, 2019 , Refereed
  • Development of a specific control system for drug logistics in disaster-hit regions, Shunji Ishiwata, Manabu Kitakoji, Tomomi Inoue, Tohru Ohtori, Takeshi Kotake, Japanese Journal of Occupational Medicine and Traumatology, Japanese Journal of Occupational Medicine and Traumatology, 66, 156 - 163, 2018 , Refereed
  • Decomposition of anthracycline anticancer drugs by ozone gas as a decontaminant, ISHIWATA Shunji, Jouranl of Medical and Hyienic Use of Ozone, Jouranl of Medical and Hyienic Use of Ozone, 25, 90 - 97, 2018 , Refereed
  • Evaluation of BLS skills by a BLS training program for pharmacy students -activation of the EMS system, rescue breathing, and AED operation, Tomomi Inoue, Shuho Yaegashi, Ryuzo Kyukawa, Shunji Ishiwata, Hiroshi Nonogi, Takeshi Kotake, Journal of Clinical Simulation Research, Journal of Clinical Simulation Research, 7, 8 - 14, 2017 , Refereed
  • Improvement and limitation of chest compression skills by a BLS training program, Tomomi Inoue, Shuho Yaegashi, Ryuzo Kyukawa, Shunji Ishiwata, Hiroshi Nonogi, Takeshi Kotake, Journal of Clinical Simulation Research, Journal of Clinical Simulation Research, 7, 19 - 24, 2017 , Refereed
  • Application of ozone gas for decontamination of nucleoside anticancer drugs, Ayumi Tsukamoto, Shunji Ishiwata, Asami Kajimoto, Ryusuke Murata, Rika Kitano, Tomomi Inoue, Takeshi Kotake, Journal of Pharmaceutical Health Care and Sciences, Journal of Pharmaceutical Health Care and Sciences, 2(26), 2016 , Refereed
  • Role of RNA-Binding Proteins in MAPK Signal Transduction Pathway, Journal of Signal Transduction, Journal of Signal Transduction, Feb. 2011
  • Production and characterization of monoclonal antibody for capture of proteins tagged with lithocholic acid., Anal. Sci., Anal. Sci., 24(11), 1475 - 1480, Nov. 2008
  • Atf1 Is a Target of the MAP Kinase Pmk1 and Regulates Cell Integrity in Fission Yeast., Mol. Biol. Cell, Mol. Biol. Cell, 18(12), 1794 - 4802, 2007
  • Pancreatic cancer stem cells: features and detection methods., Toshiyuki Ishiwata, Yoko Matsuda, Hisashi Yoshimura, Norihiko Sasaki, Shunji Ishiwata, Naoshi Ishikawa, Kaiyo Takubo, Tomio Arai, Junko Aida, Pathology oncology research : POR, Pathology oncology research : POR, 24(4), 797 - 805, Oct. 2018 , Refereed
    Summary:Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with a high incidence of distant metastasis and recurrence. Cancer stem cells (CSCs), which are pluripotent, self-renewable, and capable of forming tumors, contribute to PDAC initiation and metastasis and are responsible for resistance to chemotherapy and radiation. Three types of experimental methods are commonly used to identify CSCs: CSC-specific marker detection, a sphere-formation assay that reveals cell proliferation under non-adherent conditions, and detection of side-population (SP) cells that possess high intracellular-to-extracellular pump functions. Several CSC-specific markers have been reported in PDACs, including CD133, CD24, CD44, CXCR4, EpCAM, ABCG2, c-Met, ALDH-1, and nestin. There remains controversy regarding which markers are specific to PDAC CSCs and which are expressed alone or in combination in CSCs. Examining characteristics of isolated CSCs and discovering CSC-specific treatment options are important to improve the prognosis of PDAC cases. This review summarizes CSC-detection methods for PDAC, including CSC-marker detection, the sphere-formation assay, and detection of SP cells.
  • Role of the Small GTPase Rho3 in Golgi/Endosome Trafficking through Functional Interaction with Adaptin in Fission Yeast, Ayako Kita, Cuifang Li, Yang Yu, Nanae Umeda, Akira Doi, Mitsuko Yasuda, Shunji Ishiwata, Atsushi Taga, Yoshitaka Horiuchi, Reiko Sugiura, PLOS ONE, PLOS ONE, 6(2), Feb. 2011
    Summary:Background: We had previously identified the mutant allele of apm1(+) that encodes a homolog of the mammalian mu 1A subunit of the clathrin-associated adaptor protein-1 (AP-1) complex, and we demonstrated the role of Apm1 in Golgi/endosome trafficking, secretion, and vacuole fusion in fission yeast. Methodology/Principal Findings: In the present study, we isolated rho3+, which encodes a Rho-family small GTPase, an important regulator of exocystosis, as a multicopy-suppressor of the temperature-sensitive growth of the apm1-1 mutant cells. Overexpression of Rho3 suppressed the Cl- sensitivity and immunosuppressant sensitivity of the apm1-1 mutant cells. Overexpression of Rho3 also suppressed the fragmentation of vacuoles, and the accumulation of v-SNARE Syb1 in Golgi/endosomes and partially suppressed the defective secretion associated with apm1-deletion cells. Notably, electron microscopic observation of the rho3-deletion cells revealed the accumulation of abnormal Golgi-like structures, vacuole fragmentation, and accumulation of secretory vesicles; these phenotypes were very similar to those of the apm1-deletion cells. Furthermore, the rho3-deletion cells and apm1-deletion cells showed very similar phenotypic characteristics, including the sensitivity to the immunosuppressant FK506, the cell wall-damaging agent micafungin, Cl-, and valproic acid. Green fluorescent protein (GFP)-Rho3 was localized at Golgi/endosomes as well as the plasma membrane and division site. Finally, Rho3 was shown to form a complex with Apm1 as well as with other subunits of the clathrin-associated AP-1 complex in a GTP- and effector domain-dependent manner. Conclusions/Significance: Taken together, our findings reveal a novel role of Rho3 in the regulation of Golgi/endosome trafficking and suggest that clathrin-associated adaptor protein-1 and Rho3 co-ordinate in intracellular transport in fission yeast. To the best of our knowledge, this study provides the first evidence of a direct link between the small GTPase Rho and the clathrin-associated adaptor protein-1 in membrane trafficking.
  • In vitro assay of the interaction between Rnc1 protein and Pmp1 mRNA by affinity capillary electrophoresis with a carboxylated capillary, Atsushi Taga, Ryosuke Satoh, Shunji Ishiwata, Shuji Kodama, Atsushi Sato, Kentaro Suzuki, Reiko Sugiura, JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, 53(5), 1332 - 1337, Dec. 2010
    Summary:The interaction between Rnc1, an RNA interactive protein, and a Pmp1 mRNA was investigated by affinity capillary electrophoresis (ACE). Prior to the ACE experiments, the column performances of three capillaries (an untreated fused silica capillary, a polybrene-polyacrylic acid (PB-PAA) double layer coating capillary, and a carboxylated capillary with a covalent modification) were studied with model proteins including ribonuclease B (RNase B) and bovine serum albumin (BSA) Using an untreated fused silica and a PB-PAA double layer coating capillaries, both of the protein peaks were broad and tailing. However, using a carboxylated capillary, the protein peaks were sharp and symmetric, and migration times were repeatable (RSD < 0 4%) Further, the proteins in human serum also gave sharp peaks and its repeatability was kept at a high level by pre-treatment of a capillary inner wall with 1 M sodium chloride solution before each run. An Rnc1 protein was analyzed by ACE with background electrolytes containing various concentrations of Pmp1 sense mRNA using a carboxylated capillary. Increase in the concentration of the mRNA was found to delay the migration time of the protein. But the migration time of the protein was kept constant with increasing Pmp1 anti-sense mRNA instead of Pmp1 sense mRNA A straight line (r=0.987) was obtained by plotting 1/(migration time shift) versus 1/(Pmp1 sense mRNA concentration) and the association constant of Rnc1 protein with Pmp1 sense mRNA could be estimated to be 4 15 x 10(6) M(-1). These results suggest that the association constants of proteins with mRNAs as ligands were easily determined by the proposed method. (C) 2010 Elsevier B.V. All rights reserved.
  • The Cell Surface Protein Gene ecm33(+) Is a Target of the Two Transcription Factors Atf1 and Mbx1 and Negatively Regulates Pmk1 MAPK Cell Integrity Signaling in Fission Yeast, Hirofumi Takada, Aiko Nishida, Mitsuhiro Domae, Ayako Kita, Yuki Yamano, Atsushi Uchida, Shunji Ishiwata, Yue Fang, Xin Zhou, Takashi Masuko, Mitsuhiro Kinoshita, Kazuaki Kakehi, Reiko Sugiura, MOLECULAR BIOLOGY OF THE CELL, MOLECULAR BIOLOGY OF THE CELL, 21(4), 674 - 685, Feb. 2010
    Summary:The highly conserved fission yeast Pmk1 MAPK pathway plays a key role in cell integrity by regulating Atf1, which belongs to the ATF/cAMP-responsive element-binding (CREB) protein family. We identified and characterized ecm33(+), which encodes a glycosyl-phosphatidylinositol (GPI)-anchored cell surface protein as a transcriptional target of Pmk1 and Atf1. We demonstrated that the gene expression of Ecm33 is regulated by two transcription factors Atf1 and a MADS-box-type transcription factor Mbx1. We identified a putative ATF/CREB-binding site and an RLM1-binding site in the ecm33(+) promoter region and monitored the transcriptional activity of Atf1 or Mbx1 in living cells using a destabilized luciferase reporter gene fused to three tandem repeats of the CRE and six tandem repeats of the Rlm1-binding sequence, respectively. These reporter genes reflect the activation of the Pmk1 pathway by various stimuli, thereby enabling the real-time monitoring of the Pmk1 cell integrity pathway. Notably, the Delta ecm33 cells displayed hyperactivation of the Pmk1 signaling together with hypersensitivity to Ca2+ and an abnormal morphology, which were almost abolished by simultaneous deletion of the components of the Rho2/Pck2/Pmk1 pathway. Our results suggest that Ecm33 is involved in the negative feedback regulation of Pmk1 cell integrity signaling and is linked to cellular Ca2+ signaling.
  • Role of the RNA-binding Protein Nrd1 and Pmk1 Mitogen-activated Protein Kinase in the Regulation of Myosin mRNA Stability in Fission Yeast, Ryosuke Satoh, Takahiro Morita, Hirofumi Takada, Ayako Kita, Shunji Ishiwata, Akira Doi, Kanako Hagihara, Atsushi Taga, Yasuhiro Matsumura, Hideki Tohda, Reiko Sugiura, MOLECULAR BIOLOGY OF THE CELL, MOLECULAR BIOLOGY OF THE CELL, 20(9), 2473 - 2485, May 2009
    Summary:Myosin II is an essential component of the actomyosin contractile ring and plays a crucial role in cytokinesis by generating the forces necessary for contraction of the actomyosin ring. Cdc4 is an essential myosin II light chain in fission yeast and is required for cytokinesis. In various eukaryotes, the phosphorylation of myosin is well documented as a primary means of activating myosin II, but little is known about the regulatory mechanisms of Cdc4. Here, we isolated Nrd1, an RNA-binding protein with RNA-recognition motifs, as a multicopy suppressor of cdc4 mutants. Notably, we demonstrated that Nrd1 binds and stabilizes Cdc4 mRNA, thereby suppressing the cytokinesis defects of the cdc4 mutants. Importantly, Pmk1 mitogen-activated protein kinase (MAPK) directly phosphorylates Nrd1, thereby negatively regulating the binding activity of Nrd1 to Cdc4 mRNA. Consistently, the inactivation of Pmk1 MAPK signaling, as well as Nrd1 overexpression, stabilized the Cdc4 mRNA level, thereby suppressing the cytokinesis defects associated with the cdc4 mutants. In addition, we demonstrated the cell cycle-dependent regulation of Pmk1/Nrd1 signaling. Together, our results indicate that Nrd1 plays a role in the regulation of Cdc4 mRNA stability; moreover, our study is the first to demonstrate the posttranscriptional regulation of myosin expression by MAPK signaling.
  • Immunoprecipitation and MALDI-MS identification of lithocholic acid-tagged proteins in liver of bile duct-ligated rats, Shigeo Ikegawa, Tetsushi Yamamoto, Hiromi Ito, Shunji Ishiwata, Toshihiro Sakai, Kuniko Mitamura, Masako Maeda, JOURNAL OF LIPID RESEARCH, JOURNAL OF LIPID RESEARCH, 49(11), 2463 - 2473, Nov. 2008
    Summary:Formation of covalently bound protein adducts with lithocholic acid (LCA) might explain LCA's known carcinogenic properties and hepatotoxicity. We performed studies aimed at isolating and identifying hepatic proteins tagged with LCA, presumably via the epsilon-amino group of lysine residues. Antibodies recognizing the 3 alpha-hydroxy-5 beta-steroid moiety of LCA were generated by immunizing rabbits with immunogens in which the carboxyl group of LCA was coupled to BSA via a 6-aminohexanoic acid and/or succinic acid spacer. The resulting antibodies reacted with N-alpha (t-butoxycarbonyl)-L-lysine-epsilon-LCA, the amidated and nonamidated forms of LCA, as well as synthetically prepared LCA adducts with ovalbumin and lysozyme. Proteins tagged with LCA in the liver of bile duct-ligated rats were isolated by immunoprecipitation using these antibodies. Proteins were isolated by two-dimensional electrophoresis, and their structure was identified using matrix-assisted laser desorption ionization time-of-flight mass spectrometry and computer-assisted programs. Proteins labeled with LCA were Rab-3, Rab-12, Rab-16, and M-Ras. Rab proteins are Ras-like small GTP binding proteins that regulate vesicle trafficking pathways. The covalent binding of the Rab proteins with LCA may influence vesicular transport or binding of vesicles to their cognate membrane and may contribute to LCA-induced liver toxicity.

Books etc

  • Sourcebook of Models for Biomedical Research, Humana Press Inc., Molecular Genetic Approach to Identify Inhibitors of Signal Transduction Pathways., Joint author,   2008

Conference Activities & Talks

  • Spatial regulatory mechanism of the PI4,5P2 signaling pathway by membrane trafficking system,   2011 06
  • MAP kinase signaling regulates stress granule formation via the RNA-binding protein Nrd1,   2011 06
  • The MADS-box transcription factor Mbx1 controls Ca2+ homeostasis via the regulation of dom1+,   2011 06
  • MAP kinase signaling-dependent regulation of stress granule formation mediated by the RNA-binding protein Nrd1 in fission yeast,   2011 06
  • MADS-Box Transcription Factor Mbx1 Is Involved in the Regulation of Calcineurin Signaling through Ca2+-Homeostasis Modulation, 1st International Symposium on Carcinogenic Spiral & 9th International Conference on Protein Phospha,   2011 02 , 1st International Symposium on Carcinogenic Spiral & 9th International Conference on Protein Phospha
  • The Actin Binding Protein Cis3/PSTPIP Is Involved in the Regulation of the Pmk1 MAPK Signaling Pathway and Cytokinesis, 1st International Symposium on Carcinogenic Spiral & 9th International Conference on Protein Phospha,   2011 02 , 1st International Symposium on Carcinogenic Spiral & 9th International Conference on Protein Phospha
  • MAP Kinase Signaling Dependent Regulation of Cell Fate Mediated by the RNA-binding protein Nrd1 in Fission yeast, The 19th CDB Meeting RNA Sciences in Cell and Developmental Biology,   2010 05 , The 19th CDB Meeting RNA Sciences in Cell and Developmental Biology
  • Functional Relationship between MAP kinase Signaling and Pumilio in Fission Yeast, The 19th CDB Meeting RNA Sciences in Cell and Developmental Biology,   2010 05 , The 19th CDB Meeting RNA Sciences in Cell and Developmental Biology
  • MAPK SIGNALING-DEPENDENT REGULATION OF CELL FATE MEDIATED BY THE RNA-BINDING PROTEIN Nrd1, The 5th International Fission Yeast Meeting (Pombe 2009),   2009 10 , The 5th International Fission Yeast Meeting (Pombe 2009)
  • A NOVEL CELL SURFACE PROTEIN TPA1 ACTS AS A NEGATIVE REGULATOR OF THE PMK1 MAPK SIGNALING, The 5th International Fission Yeast Meeting (Pombe 2009),   2009 10 , The 5th International Fission Yeast Meeting (Pombe 2009)
  • The Implementation of the Model Core Curriculum toward Hospital Pharmacy Practice Experience,   2009 03
  • The regulatory mechanism of cell-fate switching by MAPK signaling and the RNA-binding protein Nrd1, 8th International Conference on Protein Phosphatases,   2008 11 , 8th International Conference on Protein Phosphatases
  • The Actin Binding Protein Cis3/PSTPIP Plays a Role in Activation of MAPK Signaling Pathways, 8th International Conference on Protein Phosphatases,   2008 11 , 8th International Conference on Protein Phosphatases
  • Cross-talk between Calcineurin Signaling and the MADS-box Transcription Factor Mbx1 in the Ca2+ Homeostasis in Fission Yeast, 8th International Conference on Protein Phosphatases,   2008 11 , 8th International Conference on Protein Phosphatases
  • The Atf1 Transcription Factor Is a Target of the Mitogen-activated Protein Kinase Pmk1 and Regulates the Expression of the tpa1+ Gene, which Encodes a Novel Cell Surface Protein in Fission Yeast, 8th International Conference on Protein Phosphatases,   2008 11 , 8th International Conference on Protein Phosphatases
  • Cbp1, an Endogenous Inhibitor of Calcineurin Signaling, Acts as a Negative Regulator of Atf1, a Downstream Transcription Factor of the Stress-activated MAP Kinase Pathway, 8th International Conference on Protein Phosphatases,   2008 11 , 8th International Conference on Protein Phosphatases
  • Observation of Interaction between RNA and GFP-Protein by Affinity Capillary Electrophoresis, 29 th International Symposium on the ?Separation of Proteins, Peptides and Polynucleotides,   2008 09 , 29 th International Symposium on the ?Separation of Proteins, Peptides and Polynucleotides
  • Study of RNA-protein interaction in MAP kinase signaling by affinity capillary electrophoresis with a carboxylated capillary, 28 th International Symposium on the ?Separation of Proteins, Peptides and Polynucleotides,   2008 09 , 28 th International Symposium on the ?Separation of Proteins, Peptides and Polynucleotides
  • The Atf1 Transcription Factor acts as a critical signaling node that links cell integrity signal and stress responses mediated by two distinct MAPK pathways., The 21st Naito Conference-Nuclear Dynamics and RNA [I],   2008 06 , The 21st Naito Conference-Nuclear Dynamics and RNA [I]
  • The Implementation of the Model Core Curriculum toward Pharmacy Practice Experience for 2-4 weeks,   2008 03
  • Development of a novel type capillary for protein analysis by capillary electrophoresis, EUROanalysis XIV,   2007 09 , EUROanalysis XIV
  • Functional interaction between Pumilio family and cell integrity signaling in fission yeast.,   2007 07
  • Apm1, theμ1 subunit of the Clathrin-Associated Adaptor Protein-1 Complex, Shares an essential function with Calcineurin in fission yeast.,   2007 07
  • The identical and phenotypic characterization of an endogenous calcineurin inhibitor Cbp1 in fission yeast.,   2007 07
  • A Survey on the Students’ Attitudes toward Pharmacy Practice Experience in Kinki University,   2007 03

Patents

  • The trainig kit and methods for anti-tumor cancer injection mixing, PCT/JP2010/66716
  • Real time assay method for MAP kinase by monitoring of mRNA and exploiting new anti-cancer drug, 特願2010-162093
  • Trainig kit and methods for anti-tumor cancer injection mixing, 特願2009-249404
  • Analysis of interaction between nucleic acid and protein by affinity capillary electrophoresis, 特願2009-207005