Histone Deacetylases and their Inhibitors as Potential Therapeutic Drugs for cholangiocarcinoma - Cell Line findings

  • Sriraksa, Ruethairat (Centre for Research and Development of medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Liver Fluke and Cholangiocarcinoma Research Center, Faculty of Medicine, khon Kaen University) ;
  • Limpaiboon, Temduang (Centre for Research and Development of medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Liver Fluke and Cholangiocarcinoma Research Center, Faculty of Medicine, khon Kaen University)
  • 발행 : 2013.04.30


Histone deacetylation mediated by histone deacetylases (HDACs) has been reported as one of the epigenetic mechanisms associated with tumorigenesis. The poor responsiveness of anticancer drugs found with cholangiocarcinoma (CCA) leads to short survival rate. We aimed to investigate mRNA expression of HDACs class I and II, and the effect of HDAC inhibitors, suberoylanilide hydroxamic acid (SAHA) and valproic acid (VPA), in CCA in vitro. Expression of HDACs was studied in CCA cell lines (M213, M214 and KKU-100) and an immortal cholangiocyte (MMNK1) by semi-quantitative reverse transcription-PCR. SAHA and VPA, as well as a classical chemotherapeutic drug 5 -fluorouacil (5-FU) were used in this study. Cell proliferation was determined by sulforhodamine assay. $IC_{50}$ and $IC_{20}$ were then analyzed for each agent and cell line. Moreover, synergistic potentional of VPA or SAHA in combination with 5-FU at sub toxic does ($IC_{20}$) of each agent was also evaluated. Statistic difference of HDACs expression or cell proliferation in each experimental condition was analyzed by Student's t-test. The result demonstrated that HDACs were expressed in all studied cell types. Both SAHA and VPA inhibited cell proliferation in a dose-dependent manner. Interestingly, KKU-100 which was less senstitive to classical chemotheraoeutic 5-FU was highly was sensitive to HDAC inhibitors. Simultaneous combination of subtoxic doses of HDAC inhibitors and 5-FU signiicantly inhibited cell proliferation in CCA cell lines compared to single sgent treatment($P{\leq}0.01$), while sequentially combined treatments were less effective. The present study showed inhibitory effects of HDACIs on cell proliferation in CCA cell lines, with synergistic antitumor potential demonstrated by simultaneous combination of VPA or SAHA with 5-FU, suggesting a novel alternative therapeutic strategy in effective treatment of CCA.


  1. Balasubramanian S, Verner, Buggy JJ (2009). Isoform-specific histone deacetylase inhibitors: the next step? Cancer Lett, 280, 211-21.
  2. Blaheta RA, Cinatl J, Jr. (2002). Anti-tumor mechanisms of valproate: a novel role for an old drug. Med Res Rev, 22, 492-511.
  3. Butler LM, Zhou X, Xu WS, et al (2002). The histone deacetylase inhibitor SAHA arrests cancer cell growth, up-regulates thioredoxin-binding protein-2, and down-regulates thioredoxin. Porc Natl Acad Sci USA, 99, 11700-5.
  4. Chen MY, Liao WS, Lu Z, et al (2011). Decitabine and suberoylanilide hydroxamic acid (SAHA) inhibit growth of ovarian cancer cell lines and xenografts while inducing expression of imprinted tumor suppressor genes, apoptosis, G2/M arrest, and autophagy. Cancer, 117, 4424-38.
  5. Cinatl J, Jr., Cinatl J, Driever PH, et al (1997). Sodium valproate inhibits in vivo growth of human neuroblastoma cells. Anticaner Drugs, 8, 958-63.
  6. Fritsche P, Seidler B, Schuler S, et al (2009). HDAC2 mediates therapeutic resistance of pancreatic cancer cells via the BH3-only protein NOXA. Gut, 58, 1399-409.
  7. Hejna M, Pruckmayer M, Raderer M (1998). The role of chemotherapy and radiation in the management of biliary cancer: a review of the literature. Eur J Cancer, 34, 977-86.
  8. Iwahashi S, Ishibashi H, Utsunomiya T, et al (2011). Effect of histone deacetylase inhibitor in combination with 5-fluorouracil on pancreas and cholangiocarcinoma cell lines. J Med Invest, 58, 106-9.
  9. Kelly WK, Marks PA (2005). Drug insight: Histone deacetylase inhibitors--development of the new targeted anticancer agent suberoylanilide hydroxamic acid. Nat Clin Pract Oncol, 2, 150-7.
  10. Khan SA, Davidson BR, Goldin R, et al (2002). Guidelines for the diagnosis and treatment of cholangiocarcinoma: consensus document. Gut, 51, 7-9.
  11. Kumagai T, Wakimoto N, Yin D, et al (2007). Histone deacetylase inhibitor, suberoylanilide hydroxamic acid (Vorinostat, SAHA) profoundly inhibits the growth of human pancreatic cancer cells. Int J Cancer, 121, 656-65.
  12. Lee MA, Woo IS, Kang JH, et al (2004). Epirubicin, cisplatin, and protracted infusion of 5-FU (ECF) in advanced intrahepatic cholangiocarcinoma. J Cancer Res Clin Oncol, 130, 346-50.
  13. Limpaiboon T (2012). Epigenetic aberrat ions in cholangiocarcinoma: potential biomarkers and promising target for novel therapeutic strategies. Asian Pac J Cancer Prev, 13, 41-5.
  14. Lin RJ, Nagy L, Inoue S, et al (1998). Role of the histone deacetylase complex in acute promyelocytic leukaemia. Nature, 391, 811-4.
  15. Marks PA, Dokmanovic M (2005). Histone deacetylase inhibitors: discovery and development as anticancer agents. Expert Opin Investing Drug, 14, 1497-511.
  16. Martin R, Jarnagin W (2003). Intrahepatic cholangiocarcinoma. Current management. Minerva Chir, 58, 469-78.
  17. Morine Y, Shimada M, Iwahashi S, et al (2012). Role of histone deacetylase expression in intrahepatic cholangiocarcinoma. Surgery, 151, 412-9.
  18. Mutze K, Langer R, Becker K, et al (2010). Histone deacetylase (HDAC) 1 and 2 expression and chemotherapy in gastric cancer. Ann Surg Oncol, 17, 3336-43.
  19. Pan LN, Lu J, Huang B (2007). HDAC inhibitors: a potential new category of anti-tumor agents. Cell Mol Immunol, 4, 337-43.
  20. Patel T (2006). Cholangiocarcinoma. Nat Clin Pract Gastroenterol Hepatol, 3, 33-42.
  21. Patra SK, Patra A, Dahiya R (2001). Histone deacetylase and DNA methyltransferase in human prostate cancer. Biochem Biophys Res Commun, 287, 705-13.
  22. Patt YZ, Hassan MM, Lozano RD, et al (2001). Phase II trial of cisplatin, interferon alpha-2b, doxorubicin, and 5-fluorouracil for biliary tract cancer. Clin Cancer Res, 7, 3375-80.
  23. Rikiishi H (2011). Autophagic and apoptotic effects of HDAC inhibitors on cancer cells. J Biomed Biotechnol, 2011, 830260.
  24. Shao Y, Gao Z, Marks PA, et al (2004). Apoptotic and autophagic cell death induced by histone deacetylase inhibitors. Porc Natl Acad Sci USA, 101, 18030-5.
  25. Sharma S, Kelly TK, Jones PA (2010). Epigenetics in cancer. Carcinogenesis, 31, 27-36.
  26. Shukla V, Vaissiere T, Herceg Z (2008). Histone acetylation and chromatin signature in stem cell identity and cancer. Mutat Res, 637, 1-15.
  27. Sia D, Tovar V, Moeini A, et al (2013). Intrahepatic cholangiocarcinoma: pathogenesis and rationale for molecular therapies. Oncogene, doi: 10.1038/onc.2012.617.
  28. Stiborova M, Eckschlager T, Poljakova J, et al (2012). The synergistic effects of DNA-targeted chemotherapeutics and histone deacetylase inhibitors as therapeutic strategies for cancer treatment. Curr Med Chem, 19, 4218-38.
  29. Thongprasert S (2005). The role of chemotherapy in cholangiocarcinoma. Ann Oncol, 16, 93-6.
  30. Venkataramani V, Rossner C, Iffland L, et al (2010). Histone deacetylase inhibitor valproic acid inhibits cancer cell proliferation via down-regulation of the alzheimer amyloid precursor protein. J Biol Chem, 285, 10678-89.
  31. Vichai V, Kirtikara K (2006). Sulforhodamine B colorimetric assay for cytotoxicity screening. Nat Protoc, 1, 1112-6.
  32. Vigushin DM, Coombes RC (2002). Histone deacetylase inhibitors in cancer treatment. Anticancer Drugs, 13, 1-13.
  33. Weichert W, Roske A, Gekeler V, et al (2008). Histone deacetylases 1, 2 and 3 are highly expressed in prostate cancer and HDAC2 expression is associated with shorter PSA relapse time after radical prostatectomy. Br J Cancer, 98, 604-10.
  34. Xia Q, Sung J, Chowdhury W, et al (2006). Chronic administration of valproic acid inhibits prostate cancer cell growth in vitro and in vivo. Cancer Res, 66, 7237-44.
  35. Yamaguchi J, Sasaki M, Sato Y, et al (2010). Histone deacetylase inhibitor (SAHA) and repression of EZH2 synergistically inhibit proliferation of gallbladder carcinoma. Cancer Sci, 101, 355-62.
  36. Zabron A, Edwards RJ, Khan SA (2013). The challenge of cholangiocarcinoma: dissecting the molecular mechanisms of an insidious cancer. Dis Model Mech, 6, 281-92.

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