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Tumor Biology

, Volume 35, Issue 11, pp 11523–11532 | Cite as

Down-regulation of HDAC5 inhibits growth of human hepatocellular carcinoma by induction of apoptosis and cell cycle arrest

  • Jian Fan
  • Bin Lou
  • Wei Chen
  • Jie Zhang
  • Sha Lin
  • Fei-fei Lv
  • Yu Chen
Research Article

Abstract

Histone deacetylases (HDACs) play a critical role in the proliferation, differentiation, and apoptosis of cancer cells. An obstacle for the application of HDAC inhibitors as effective anti-cancer therapeutics is that our current knowledge on the contributions of different HDACs in various cancer types remains scarce. The present study reported that the mRNA and protein levels of HDAC5 were up-regulated in human hepatocellular carcinoma (HCC) tissues and cells as shown by quantitative real-time PCR and Western blot. MTT assay and BrdU incorporation assay showed that the down-regulation of HDAC5 inhibited cell proliferation in HepG2, Hep3B, and Huh7 cell lines. Data from in vivo xenograft tumorigenesis model also demonstrated the anti-proliferative effect of HDAC5 depletion on tumor cell growth. Furthermore, the suppression of HDAC5 promoted cell apoptosis and induced G1-phase cell cycle arrest in HCC cells. On the molecular level, we observed altered expression of apoptosis-related proteins such as p53, bax, bcl-2, cyto C, and caspase 3 in HDAC5-shRNA-transfected cells. Knockdown of HDAC5 led to a significant up-regulation of p21 and down-regulation of cyclin D1 and CDK2/4/6. We also found that the down-regulation of HDAC5 substantially increased p53 stability and promoted its nuclear localization and transcriptional activity. Our study suggested that knockdown of HDAC5 could inhibit cancer cell proliferation by the induction of cell cycle arrest and apoptosis; thus, suppression of HDAC5 may be a viable option for treating HCC patients.

Keywords

Hepatocellular carcinoma Cell proliferation HDAC5 shRNA 

Notes

Acknowledgments

This research was supported by the State Major Science and Technology Special Projects during the period of China State 12th 5-year plan (grant 2012ZX10002).

Conflicts of interest

None

References

  1. 1.
    El-Serag HB. Hepatocellular carcinoma. N Engl J Med. 2011;365:1118–27.PubMedCrossRefGoogle Scholar
  2. 2.
    Yang Y, Jin C, Li H, et al. Improved radiosensitizing effect of the combination of etanidazole and paclitaxel for hepatocellular carcinoma in vivo. Exp Ther Med. 2012;3:299–303.PubMedCentralPubMedGoogle Scholar
  3. 3.
    Zhang C, Ling Y, Zhang C, et al. The silencing of reck gene is associated with promoter hypermethylation and poor survival in hepatocellular carcinoma. Int J Biol Sci. 2012;8:451–8.PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Minagawa M, Makuuchi M, Takayama T, et al. Selection criteria for repeat hepatectomy in patients with recurrent hepatocellular carcinoma. Ann Surg. 2003;238:703–10.PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Johnson FM, Saigal B, Talpaz M, et al. Dasatinib (BMS-354825) tyrosine kinase inhibitor suppresses invasion and induces cell cycle arrest and apoptosis of head and neck squamous cell carcinoma and non-small cell lung cancer cells. Clin Cancer Res. 2005;11:6924–32.PubMedCrossRefGoogle Scholar
  6. 6.
    Yan K, Zhang C, Feng J, et al. Induction of G1 cell cycle arrest and apoptosis by berberine in bladder cancer cells. Eur J Pharmacol. 2011;661:1–7.PubMedCrossRefGoogle Scholar
  7. 7.
    West AC, Mattarollo SR, Shortt J, et al. An intact immune system is required for the anticancer activities of histone deacetylase inhibitors. Cancer Res. 2013;73:7265–76.PubMedCrossRefGoogle Scholar
  8. 8.
    Sebastian C, Zwaans BM, Silberman DM, et al. The histone deacetylase SIRT6 is a tumor suppressor that controls cancer metabolism. Cell. 2012;151:1185–99.PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Mottet D, Pirotte S, Lamour V, et al. HDAC4 represses p21(WAF1/Cip1) expression in human cancer cells through a Sp1-dependent, p53-independent mechanism. Oncogene. 2009;28:243–56.PubMedCrossRefGoogle Scholar
  10. 10.
    Wilson AJ, Byun DS, Nasser S, et al. HDAC4 promotes growth of colon cancer cells via repression of p21. Mol Biol Cell. 2008;19:4062–75.PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Zhu C, Chen Q, Xie Z, et al. The role of histone deacetylase 7 (HDAC7) in cancer cell proliferation: regulation on c-Myc. J Mol Med (Berl). 2011;89:279–89.CrossRefGoogle Scholar
  12. 12.
    Wagner JM, Hackanson B, Lubbert M, et al. Histone deacetylase (HDAC) inhibitors in recent clinical trials for cancer therapy. Clin Epigenetics. 2010;1:117–36.PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Zhang Y, Matkovich SJ, Duan X, et al. Receptor-independent protein kinase c alpha (PKCalpha) signaling by calpain-generated free catalytic domains induces HDAC5 nuclear export and regulates cardiac transcription. J Biol Chem. 2011;286:26943–51.PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Osada H, Tatematsu Y, Saito H, et al. Reduced expression of class ii histone deacetylase genes is associated with poor prognosis in lung cancer patients. Int J Cancer. 2004;112:26–32.PubMedCrossRefGoogle Scholar
  15. 15.
    Milde T, Oehme I, Korshunov A, et al. HDAC5 and HDAC9 in medulloblastoma: novel markers for risk stratification and role in tumor cell growth. Clin Cancer Res. 2010;16:3240–52.PubMedCrossRefGoogle Scholar
  16. 16.
    Lachenmayer A, Toffanin S, Cabellos L, et al. Combination therapy for hepatocellular carcinoma: additive preclinical efficacy of the hdac inhibitor panobinostat with sorafenib. J Hepatol. 2012;56:1343–50.PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Liu H, Li P, Zhai Y, et al. Diagnostic value of glypican-3 in serum and liver for primary hepatocellular carcinoma. World J Gastroenterol. 2010;16:4410–5.PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    Glaser KB, Li J, Staver MJ, et al. Role of class I and class II histone deacetylases in carcinoma cells using siRNA. Biochem Biophys Res Commun. 2003;310:529–36.PubMedCrossRefGoogle Scholar
  19. 19.
    Valenzuela-Fernandez A, Cabrero JR, Serrador JM, et al. HDAC6: a key regulator of cytoskeleton, cell migration and cell-cell interactions. Trends Cell Biol. 2008;18:291–7.PubMedCrossRefGoogle Scholar
  20. 20.
    Lee CK, Wang S, Huang X, et al. HDAC inhibition synergistically enhances alkylator-induced dna damage responses and apoptosis in multiple myeloma cells. Cancer Lett. 2010;296:233–40.PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Peixoto P, Castronovo V, Matheus N, et al. HDAC5 is required for maintenance of pericentric heterochromatin, and controls cell-cycle progression and survival of human cancer cells. Cell Death Differ. 2012;19:1239–52.PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    La Vignera S, Condorelli R, Vicari E, et al. Effects of varicocelectomy on sperm DNA fragmentation, mitochondrial function, chromatin condensation, and apoptosis. J Androl. 2012;33:389–96.PubMedCrossRefGoogle Scholar
  23. 23.
    Ren W, Beebe SJ. An apoptosis targeted stimulus with nanosecond pulsed electric fields (nsPEFs) in E4 squamous cell carcinoma. Apoptosis. 2011;16:382–93.PubMedCentralPubMedCrossRefGoogle Scholar
  24. 24.
    Weinert T, Hopper AK. tRNA traffic meets a cell-cycle checkpoint. Cell. 2007;131:838–40.PubMedCrossRefGoogle Scholar
  25. 25.
    Bai Y, Mao QQ, Qin J, et al. Resveratrol induces apoptosis and cell cycle arrest of human T24 bladder cancer cells in vitro and inhibits tumor growth in vivo. Cancer Sci. 2010;101:488–93.PubMedCrossRefGoogle Scholar
  26. 26.
    Yang L, Besschetnova TY, Brooks CR, et al. Epithelial cell cycle arrest in G2/M mediates kidney fibrosis after injury. Nat Med. 2010;16(535–43):1–143.Google Scholar
  27. 27.
    McKinsey TA, Zhang CL, Lu J, et al. Signal-dependent nuclear export of a histone deacetylase regulates muscle differentiation. Nature. 2000;408:106–11.PubMedCrossRefGoogle Scholar
  28. 28.
    Ropero S, Esteller M. The role of histone deacetylases (HDACs) in human cancer. Mol Oncol. 2007;1:19–25.PubMedCrossRefGoogle Scholar
  29. 29.
    Richon VM, Sandhoff TW, Rifkind RA, et al. Histone deacetylase inhibitor selectively induces p21WAF1 expression and gene-associated histone acetylation. Proc Natl Acad Sci U S A. 2000;97:10014–9.PubMedCentralPubMedCrossRefGoogle Scholar
  30. 30.
    Zhao Y, Tan J, Zhuang L, et al. Inhibitors of histone deacetylases target the Rb-E2F1 pathway for apoptosis induction through activation of proapoptotic protein Bim. Proc Natl Acad Sci U S A. 2005;102:16090–5.PubMedCentralPubMedCrossRefGoogle Scholar
  31. 31.
    Kramer OH, Gottlicher M, Heinzel T. Histone deacetylase as a therapeutic target. Trends Endocrinol Metab. 2001;12:294–300.PubMedCrossRefGoogle Scholar
  32. 32.
    Marks PA, Richon VM, Breslow R, et al. Histone deacetylase inhibitors as new cancer drugs. Curr Opin Oncol. 2001;13:477–83.PubMedCrossRefGoogle Scholar
  33. 33.
    Hu W, Ge Y, Ojcius DM, et al. P53 signalling controls cell cycle arrest and caspase-independent apoptosis in macrophages infected with pathogenic leptospira species. Cell Microbiol. 2013;15:1642–59.PubMedGoogle Scholar
  34. 34.
    Qiao D, Meyer K, Friedl A. Glypican-1 stimulates Skp2 autoinduction loop and G1/S transition in endothelial cells. J Biol Chem. 2012;287:5898–909.PubMedCentralPubMedCrossRefGoogle Scholar
  35. 35.
    Abe K, Naruse C, Kato T, et al. Loss of heterochromatin protein 1 gamma reduces the number of primordial germ cells via impaired cell cycle progression in mice. Biol Reprod. 2011;85:1013–24.PubMedCrossRefGoogle Scholar
  36. 36.
    Jin L, Li C, Xu Y, et al. Epigallocatechin gallate promotes p53 accumulation and activity via the inhibition of MDM2-mediated p53 ubiquitination in human lung cancer cells. Oncol Rep. 2013;29:1983–90.PubMedGoogle Scholar
  37. 37.
    Yuan J, Luo K, Zhang L, et al. Usp10 regulates p53 localization and stability by deubiquitinating p53. Cell. 2010;140:384–96.PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Jian Fan
    • 1
  • Bin Lou
    • 1
  • Wei Chen
    • 2
  • Jie Zhang
    • 1
  • Sha Lin
    • 1
  • Fei-fei Lv
    • 1
  • Yu Chen
    • 1
  1. 1.Department of Laboratory Medicine, First Affiliated Hospital, School of MedicineZhejiang UniversityHangzhouChina
  2. 2.Department of Hepatiobiliary and Pancreatic surgery, Second Affiliated Hospital, School of MedicineZhejiang UniversityHangzhouChina

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