title: The Rho kinase inhibitor fasudil is involved in p53-mediated apoptosis in human hepatocellular carcinoma cells.
abstract: OBJECTIVE
  Rho kinase is an important factor in tumor progression. We demonstrated that Rho kinase-associated coil-containing protein kinase (ROCK) is expressed in hepatic tissues in hepatocellular carcinoma (HCC) and confirmed its roles in cell survival in HCC cells using the ROCK inhibitor, fasudil.
  
  
  METHODS
  ROCK protein levels were estimated in hepatic tissues with HCC compared with healthy liver tissues or hepatic hemangioma tissues using immunohistochemistry. Furthermore, HepG2 and Huh7 cells were cultured with ROCK inhibitor, fasudil for 24 h in vitro. Cell proliferation was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt assay, and apoptotic cells were detected by cell death ELISA. The expression apoptosis-related proteins were analyzed using Western blotting.
  
  
  RESULTS
  Fasudil significantly decreased cell proliferation and induced apoptosis mediated by increases in p53, Bax, caspase-9, and caspase-3 in HepG2 and Huh7 cells. The induction of apoptosis was inhibited in HCC cells precultured with p53 decoy oligodeoxynucleotide.
  
  
  CONCLUSIONS
  These results suggest that ROCK inhibits the p53-mediated apoptosis pathway in HCC. Fasudil may thus be a beneficial approach to HCC therapy.
authors.affiliation: Department of Pharmacology, St. Marianna University School of Medicine, 2-16-1, Sugao, Miyamae-ku, Kawasaki, Kanagawa, 216-8511, Japan. takebay@marianna-u.ac.jp
authors.forename: Yuko
authors.initials: Y
authors.lastname: Takeba
authors.forename: Naoki
authors.initials: N
authors.lastname: Matsumoto
authors.forename: Minoru
authors.initials: M
authors.lastname: Watanabe
authors.forename: Sachiko
authors.initials: S
authors.lastname: Takenoshita-Nakaya
authors.forename: Yuki
authors.initials: Y
authors.lastname: Ohta
authors.forename: Toshio
authors.initials: T
authors.lastname: Kumai
authors.forename: Masayuki
authors.initials: M
authors.lastname: Takagi
authors.forename: Satoshi
authors.initials: S
authors.lastname: Koizumi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Asakura
authors.forename: Takehito
authors.initials: T
authors.lastname: Otsubo
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D008099:Liver / Q000201:enzymology; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000473:pathology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*; D054460:rho-Associated Kinases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000502:physiology
chemical_list: D047428:Protein Kinase Inhibitors; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D054460:rho-Associated Kinases; C049347:fasudil