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Knockdown of PPP5C Inhibits Growth of Hepatocellular Carcinoma Cells In Vitro

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Abstract

Ser/Thr protein phosphatase 5 (PPP5C) has been reported to participate in tumor progression. However, its functional role in hepatocellular carcinoma (HCC) remains unknown yet. In this study, we firstly evaluated the expression levels of PPP5C in six HCC cell lines by real-time PCR and found that PPP5C was widely expressed in HCC cells. To explore the role of PPP5C in HCC cell growth, lentivirus-mediated short hairpin RNA (shRNA) was employed to silence PPP5C expression in HepG2 and Bel-7404 cells. The expression of PPP5C was significantly downregulated in PPP5C knockdown cells. Knockdown of PPP5C markedly suppressed the proliferation and colony formation ability of HCC cells. Moreover, cell cycle analysis showed that PPP5C depletion in HepG2 cells led to G0/G1 phase and G2/M phase arrest. We demonstrate for the first time that PPP5C is essential for growth of HCC cells, which suggests that inhibition of PPP5C by RNAi may be a potential therapeutic strategy for the treatment of HCC.

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References

  1. Rahman, R., Hammoud, G. M., Almashhrawi, A. A., Ahmed, K. T., & Ibdah, J. A. (2013). Primary hepatocellular carcinoma and metabolic syndrome: an update. World Journal of Gastrointestinal Oncology, 5, 186–194.

    Article  Google Scholar 

  2. Bosch, F. X., Ribes, J., Diaz, M., & Cleries, R. (2004). Primary liver cancer: worldwide incidence and trends. Gastroenterology, 127, S5–S16.

    Article  Google Scholar 

  3. Wang, Z. G., Zhang, G. F., Wu, J. C., & Jia, M. K. (2013). Adjuvant therapy for hepatocellular carcinoma: current situation and prospect. Drug Discoveries and Therapeutics, 7, 137–143.

    CAS  Google Scholar 

  4. Agnello, F., Salvaggio, G., Cabibbo, G., Maida, M., Lagalla, R., Midiri, M., & Brancatelli, G. (2013). Imaging appearance of treated hepatocellular carcinoma. World Journal of Hepatology, 5, 417–424.

    Article  Google Scholar 

  5. Mumby, M. C., & Walter, G. (1993). Protein serine/threonine phosphatases: structure, regulation, and functions in cell growth. Physiological Reviews, 73, 673–699.

    CAS  Google Scholar 

  6. Cohen, P. T., Brewis, N. D., Hughes, V., & Mann, D. J. (1990). Protein serine/threonine phosphatases; an expanding family. FEBS Letters, 268, 355–359.

    Article  CAS  Google Scholar 

  7. Cohen, P. T. (1997). Novel protein serine/threonine phosphatases: variety is the spice of life. Trends in Biochemical Sciences, 22, 245–251.

    Article  CAS  Google Scholar 

  8. Zuo, Z., Dean, N. M., & Honkanen, R. E. (1998). Serine/threonine protein phosphatase type 5 acts upstream of p53 to regulate the induction of p21(WAF1/Cip1) and mediate growth arrest. Journal of Biological Chemistry, 273, 12250–12258.

    Article  CAS  Google Scholar 

  9. Golden, T., Swingle, M., & Honkanen, R. E. (2008). The role of serine/threonine protein phosphatase type 5 (PP5) in the regulation of stress-induced signaling networks and cancer. Cancer Metastasis Reviews, 27, 169–178.

    Article  CAS  Google Scholar 

  10. Swingle, M. R., Honkanen, R. E., & Ciszak, E. M. (2004). Structural basis for the catalytic activity of human serine/threonine protein phosphatase-5. Journal of Biological Chemistry, 279, 33992–33999.

    Article  CAS  Google Scholar 

  11. Hinds, T. D., Jr., & Sanchez, E. R. (2008). Protein phosphatase 5. International Journal of Biochemistry and Cell Biology, 40, 2358–2362.

    Article  CAS  Google Scholar 

  12. Yang, J., Roe, S. M., Cliff, M. J., Williams, M. A., Ladbury, J. E., Cohen, P. T., & Barford, D. (2005). Molecular basis for TPR domain-mediated regulation of protein phosphatase 5. EMBO Journal, 24, 1–10.

    Article  Google Scholar 

  13. Golden, T., Aragon, I. V., Zhou, G., Cooper, S. R., Dean, N. M., & Honkanen, R. E. (2004). Constitutive over expression of serine/threonine protein phosphatase 5 (PP5) augments estrogen-dependent tumor growth in mice. Cancer Letters, 215, 95–100.

    Article  CAS  Google Scholar 

  14. Golden, T., Aragon, I. V., Rutland, B., Tucker, J. A., Shevde, L. A., Samant, R. S., Zhou, G., Amable, L., Skarra, D., & Honkanen, R. E. (2008). Elevated levels of Ser/Thr protein phosphatase 5 (PP5) in human breast cancer. Biochimica et Biophysica Acta, 1782, 259–270.

    Article  CAS  Google Scholar 

  15. Urban, G., Golden, T., Aragon, I. V., Cowsert, L., Cooper, S. R., Dean, N. M., & Honkanen, R. E. (2003). Identification of a functional link for the p53 tumor suppressor protein in dexamethasone-induced growth suppression. Journal of Biological Chemistry, 278, 9747–9753.

    Article  CAS  Google Scholar 

  16. Urban, G., Golden, T., Aragon, I. V., Scammell, J. G., Dean, N. M., & Honkanen, R. E. (2001). Identification of an estrogen-inducible phosphatase (PP5) that converts MCF-7 human breast carcinoma cells into an estrogen-independent phenotype when expressed constitutively. Journal of Biological Chemistry, 276, 27638–27646.

    Article  CAS  Google Scholar 

  17. Morita, K., Saitoh, M., Tobiume, K., Matsuura, H., Enomoto, S., Nishitoh, H., & Ichijo, H. (2001). Negative feedback regulation of ASK1 by protein phosphatase 5 (PP5) in response to oxidative stress. EMBO Journal, 20, 6028–6036.

    Article  CAS  Google Scholar 

  18. Wechsler, T., Chen, B. P., Harper, R., Morotomi-Yano, K., Huang, B. C., Meek, K., Cleaver, J. E., Chen, D. J., & Wabl, M. (2004). DNA-PKcs function regulated specifically by protein phosphatase 5. Proceedings of the National Academy of Sciences of the United States of America, 101, 1247–1252.

    Article  CAS  Google Scholar 

  19. Amable, L., Grankvist, N., Largen, J. W., Ortsater, H., Sjoholm, A., & Honkanen, R. E. (2011). Disruption of serine/threonine protein phosphatase 5 (PP5:PPP5c) in mice reveals a novel role for PP5 in the regulation of ultraviolet light-induced phosphorylation of serine/threonine protein kinase Chk1 (CHEK1). Journal of Biological Chemistry, 286, 40413–40422.

    Article  CAS  Google Scholar 

  20. Yamaguchi, Y., Katoh, H., Mori, K., & Negishi, M. (2002). Galpha(12) and Galpha(13) interact with Ser/Thr protein phosphatase type 5 and stimulate its phosphatase activity. Current Biology, 12, 1353–1358.

    Article  CAS  Google Scholar 

  21. Chen, M. S., Silverstein, A. M., Pratt, W. B., & Chinkers, M. (1996). The tetratricopeptide repeat domain of protein phosphatase 5 mediates binding to glucocorticoid receptor heterocomplexes and acts as a dominant negative mutant. Journal of Biological Chemistry, 271, 32315–32320.

    Article  CAS  Google Scholar 

  22. Ali, A., Zhang, J., Bao, S., Liu, I., Otterness, D., Dean, N. M., Abraham, R. T., & Wang, X. F. (2004). Requirement of protein phosphatase 5 in DNA-damage-induced ATM activation. Genes and Development, 18, 249–254.

    Article  CAS  Google Scholar 

  23. Zhang, J., Bao, S., Furumai, R., Kucera, K. S., Ali, A., Dean, N. M., & Wang, X. F. (2005). Protein phosphatase 5 is required for ATR-mediated checkpoint activation. Molecular and Cellular Biology, 25, 9910–9919.

    Article  CAS  Google Scholar 

  24. Jeong, J. Y., Johns, J., Sinclair, C., Park, J. M., & Rossie, S. (2003). Characterization of Saccharomyces cerevisiae protein Ser/Thr phosphatase T1 and comparison to its mammalian homolog PP5. BMC Cell Biology, 4, 3.

    Article  Google Scholar 

  25. Zhou, G., Golden, T., Aragon, I. V., & Honkanen, R. E. (2004). Ser/Thr protein phosphatase 5 inactivates hypoxia-induced activation of an apoptosis signal-regulating kinase 1/MKK-4/JNK signaling cascade. Journal of Biological Chemistry, 279, 46595–46605.

    Article  CAS  Google Scholar 

  26. Chebotaev, D., Yemelyanov, A., & Budunova, I. (2007). The mechanisms of tumor suppressor effect of glucocorticoid receptor in skin. Molecular Carcinogenesis, 46, 732–740.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of General Surgery, The Second Affiliated Hospital of Harbin Medical University, No. 148 Baojian Road, Nangang District, Harbin, 150086, China

    Liang Feng, Peng Sun, Zhiyu Li & Shibo Sun

  2. Department of General Surgery, The Fourth Affiliated Hospital of Harbin Medical University, No. 31 Yinghang Road, Nangang District, Harbin, 150001, China

    Ming Liu

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  1. Liang Feng
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  2. Peng Sun
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Correspondence to Ming Liu or Shibo Sun.

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Feng, L., Sun, P., Li, Z. et al. Knockdown of PPP5C Inhibits Growth of Hepatocellular Carcinoma Cells In Vitro. Appl Biochem Biotechnol 175, 526–534 (2015). https://doi.org/10.1007/s12010-014-1281-8

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  • DOI: https://doi.org/10.1007/s12010-014-1281-8

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