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The Rho kinase inhibitor fasudil is involved in p53-mediated apoptosis in human hepatocellular carcinoma cells

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Abstract

Purpose

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.

Conclusion

These results suggest that ROCK inhibits the p53-mediated apoptosis pathway in HCC. Fasudil may thus be a beneficial approach to HCC therapy.

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Acknowledgments

This work was supported in the part by grants under the Research on Publicly Essential Drugs and Medical Devices of Health and Labor Sciences Research from the Ministry of Health, Labor and Welfare of Japan. Furthermore, we thank Ms. Shigeko Onuma of the Department of Pathology, St. Marianna University School of Medicine for technical support.

Conflict of interest

There is no conflict of interest involved in this manuscript.

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Authors and Affiliations

  1. Department of Pharmacology, St. Marianna University School of Medicine, 2-16-1, Sugao, Miyamae-ku, Kawasaki, Kanagawa, 216-8511, Japan

    Yuko Takeba, Naoki Matsumoto, Sachiko Takenoshita-Nakaya & Yuki Ohta

  2. Institute of Experimental Animals, St. Marianna Graduate School of Medicine, Miyamae-ku, Kawasaki, Kanagawa, 216-8511, Japan

    Minoru Watanabe

  3. Department of Pharmacogenetics, St. Marianna Graduate School of Medicine, Miyamae-ku, Kawasaki, Kanagawa, 216-8511, Japan

    Toshio Kumai

  4. Department of Pathology, St. Marianna University School of Medicine, 2-16-1, Sugao, Miyamae-ku, Kawasaki, Kanagawa, 216-8511, Japan

    Masayuki Takagi

  5. Department of Surgery, St. Marianna University School of Medicine, 2-16-1, Sugao, Miyamae-ku, Kawasaki, Kanagawa, 216-8511, Japan

    Satoshi Koizumi, Takeshi Asakura & Takehito Otsubo

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  1. Yuko Takeba
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  2. Naoki Matsumoto
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Correspondence to Yuko Takeba.

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Takeba, Y., Matsumoto, N., Watanabe, M. et al. The Rho kinase inhibitor fasudil is involved in p53-mediated apoptosis in human hepatocellular carcinoma cells. Cancer Chemother Pharmacol 69, 1545–1555 (2012). https://doi.org/10.1007/s00280-012-1862-6

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  • DOI: https://doi.org/10.1007/s00280-012-1862-6

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