Summary
Although interferon-α (IFN-α) has shown great promise in the treatment of chronic viral hepatitis, the anti-tumour effect of this agent in the therapy of liver cancer is unclear. Recent studies have demonstrated that differentiation-inducing agents could modulate the responsiveness of cancer cells to IFN-α by regulating the expression of signal transducers and activators of transcription (STAT) proteins, a group of transcription factors which play important roles in the IFN signalling pathway. We have reported that sodium butyrate is a potent differentiation inducer for human hepatoma cells. In this study, we investigated whether this drug could regulate the expression of STAT proteins and enhance the anti-tumour effect of IFN-α in hepatoma cells. We found that sodium butyrate specifically activated STAT1 gene expression and enhanced IFN-α-induced phosphorylation and activation of STAT1 proteins. Co-treatment with these two drugs led to G1 growth arrest, accompanied by down-regulation of cyclin D1 and up-regulation of p21WAF-1, and accumulation of hypophosphorylated retinoblastoma protein in hepatoma cells. Additionally, internucleosomal DNA fragmentation, a biological hallmark of apoptosis, was detected in hepatoma cells after continuous incubation with a combination of these two drugs for 72 h. Our results show that sodium butyrate potently enhances the anti-tumour effect of IFN-α in vitro and suggest that a rational combination of these two drugs may be useful for the treatment of liver cancer.
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Alexander, G. J., Brahm, J., Fagan, E. A., Smith, H. M., Daniels, H. M., Eddleston, A. L. & Williams, R. (1987). Loss of HBsAg with interferon therapy in chronic hepatitis virus infection. Lancet 2: 66–69.
Boue, F., Pastran, Z., Spielmann, M., Chevalier, T. L., Subirana, R., Sevin, D., Paoletti, C., Brandely, M., Avril, M. F., Sancho-Garnier, H. & Tursz, T. (1990). A phase I trial with recombinant interferon γ (Rousell UCLAF) in advanced cancer patients. Cancer Immunol Immunother 32: 67–70.
Chin, Y. E., Kitagawa, M., Su, W. C. S., You, Z. H., Iwamoto, Y. & Fu, X. Y. (1996). Cell growth arrest and induction of cyclin-dependent kinase inhibitor p21WAF-1/CIP1 mediated by STAT1. Science 272: 719–722.
Chin, Y. E., Kitagawa, M., Kuida, K., Flavell, R. A. & Fu, X. Y. (1997). Activation of the STAT signaling pathway can cause expression of caspase 1 and apoptosis. Mol Cell Biol 17: 5328–5337.
Darnell, J. E. Jr (1997). STATs and gene regulation. Science 277: 1630–1635.
Decker, T., Lew, D. J. & Darnell, J. E. Jr (1991). Two distinct alpha-interferon dependent signal transduction pathways may contribute to activation of transcription to the guanylate-binding protein gene. Mol Cell Biol 13: 5147–5153.
El-Deiry, W., Tokino, T., Velculescu, V. E., Levy, D. B., Parsons, R., Trent, J. M., Lin, D., Mercer, W. E., Kinzler, K. W. & Vogelstein, B. (1993). WAF-1, a potential mediator of p53 tumor suppression. Cell 75: 817–825.
El-Deiry, W., Harper, J. W., O’Connor, P. M., Velculescu, V. E., Canman, C. E., Jackman, J., Pietenpol, J. A., Burrell, M., Hill, D. E., Wang, Y., Wiman, K. G., Mercer, W. E., Kastan, M. B., Kohn, K. W., Elledge, S. J., Kinzler, K. W. & Vogelstein, B. (1994). WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res 54: 1169–1174.
Foster, G. R., Ackrill, A. M., Goldin, R. D., Kerr, I. M., Thomas, H. C. & Stark, G. R. (1991). Expression of the terminal protein region of hepatitis B virus inhibits cellular responses to interferon α and γ double-stranded RNA. Proc Natl Acad Sci USA 88: 2888–2892.
Goto, I., Yamamoto-Yamagichi, Y. & Honma, Y. (1996). Enhancement of sensitivity of human lung adenocarcinoma cells to growth-inhibitory activity of interferon α by differentiation-inducing agents. Br J Cancer 74: 546–554.
Gutterman, J. U. (1994). Cytokine therapeutics: lessons from interferon α. Proc Natl Acad Sci USA 91: 1198–1205.
Harper, J. W., Adami, G. R., Wei, N., Keyomarsi, K. & Elledge, S. J. (1993). The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell 75: 805–816.
Harper, J. W., Elledge, S. J., Keyomarsi, K., Dynlacht, B., Tsai, L. H., Zhang, P., Dobrowolski, S., Bai, C., Connell-Crowley, L., Swindell, E., Fox, M. P. & Wei, N. (1995). Inhibition of cyclin-dependent kinases by p21. Mol Biol Cell 6: 387–400.
Hung, W. C. & Chuang, L. Y. (1996). Induction of apoptosis by sphingosine-1-phosphate in human hepatoma cells is associated with enhanced expression of bax gene product. Biochem Biophys Res Commun 229: 11–15.
Hung, W. C., Yang, M. L., Chang, C. C., Tsai, J. H. & Chuang, L. Y. (1995). Differential regulation of EGF production, EGF receptor binding, and cellular growth by sodium butyrate in Hep3B and PLC/PRF/5 human hepatoma cells. Inter J Oncol 7: 1089–1093.
Kolla, V., Lindner, D. J., Weihua, X., Borden, E. C. & Kalvakolanu, D. V. (1996). Modulation of interferon-inducible gene expression by retinoic acid: up-regulation of STAT1 protein in interferon-unresponsive cells. J Biol Chem 271: 10508–10514.
Korenman, J., Baker, B., Waggoner, J., Everhart, J. E., Di-Bisceglie, A. M. & Hoofnagle, J. H. (1991). Long-term remission of chronic hepatitis B after alpha-interferon therapy. Ann Intern Med 114: 629–634.
Kumar, S. (1995). ICE-like proteases in apoptosis. Trends Biochem Sci 20: 198–202.
Lai, C. L., Wu, P. C., Lok, A. S. F., Lin, H. J., Ngan, H., Lau, J. Y. N., Chung, H. T., Ng, M. M. T., Yeoh, E. K. & Arnold, M. (1989). Recombinant interferon α2 is superior to doxorubin for inoperable hepatocellular carcinoma: a prospective randomised trial. Br J Cancer 60: 928–933.
Lee, C. K., Bluyssen, H. A. R. & Levy, D. E. (1997). Regulation of interferon-α responsiveness by the duration of janus kinase activity. J Biol Chem 272: 21872–21877.
Lippman, S. M., Parkinson, D. R., Itri, L. M., Weber, R. S., Schantz, S. P., Ota, D. M., Schusterman, M. A., Krakoff, I. H., Gutterman, J. U. & Hong, W. K. (1992a). 13-cis-retinoic acid and interferon α-2a: effective combination therapy for advanced squamous cell carcinoma of the skin. J Natl Cancer Inst 84: 235–241.
Lippman, S. M., Kavanagh, J. J., Pareds-Espinoza, M., Delgadillo-Madrueno, P., Paredes-Casillas, P., Hong, W. K., Holdener, E. & Krakoff, I. H. (1992b). 13-cis-retinoic acid plus interferon α-2a: highly active systemic therapy for squamous cell carcinoma of the cervix. J Natl Cancer Inst 84: 241–245.
Muller, M., Laxton, C., Briscoe, J., Schindler, C., Improta, T., Darnell, J. E. Jr, Stark, G. R. & Kerr, I. M. (1993). Complementation of a mutant cell line: central role of the 91 kDa polypeptide of ISGF3 in the interferon-alpha and -gamma signal transduction pathways. EMBO J 12: 4221–4228.
Nair, P. V., Tong, M. J., Kempf, R., Co, R., Lee, S. D. & Venturi, C. L. (1985). Clinical serologic and immunological effects of human leukocyte interferon in HBsAg-positive primary hepatocellular carcinoma. Cancer 56: 1018–1023.
Nakano, K., Mizuno, T., Sowa, Y., Orita, T., Yoshino, T., Okuyam, Y., Fujita, T., Ohtani-Fujita, N., Matsukawa, Y., Tokino, T., Yamagishi, H., Oka, T., Nomura, H. & Sakai, T. (1997). Butyrate activates the WAF1/CIP1 gene promoter through Sp1 sites in a p53-negative human colon cancer cell line. J Biol Chem 272: 22199–22206.
Pelicano, L., Li, F., Schindler, C. & Chelbi-Alix, M. K. (1997). Retinoic acid enhances the expression of interferon-induced proteins: evidence for multiple mechanisms of action. Oncogene 15: 2349–2359.
Pellegrini, S. & Dusanter-Fourt, I. (1997). The structure, regulation and function of the Janus kinases (JAKs) and the signal transducers and activators of transcription (STATs). Eur J Biochem 248: 615–633.
Sachs, E., Di-Biscegle, A. M., Dusheiko, G. M., Song, E., Lyons, S. F., Schoub, B. D. & Kew, M. C. (1985). Treatment of hepatocellular carcinoma with recombinant leucocyte interferon: a pilot study. Br J Cancer 52: 105–109.
Tur-Kaspa, R., Teicher, L., Laub, O., Itin, A., Dagan D Bloom, B. R. & Shafritz, D. A. (1990). Alpha interferon suppresses hepatitis B virus enhancer activity and reduces viral gene transcription. Virol 64: 1821–1824.
Weihua, X., Kolla, V. & Kalvakolanu, D. V. (1997). Modulation of interferon action by retinoids: induction of murine STAT1 gene expression by retinoic acid. J Biol Chem 272: 9742–9748.
Xiong, Y., Hannon, G. J., Zhang, H., Casso, D., Kobayashi, R. & Beach, D. (1993). p21 is a universal inhibitor of cyclin kinases. Nature 366: 701–704.
Xu, B., Grander, D., Sangfelt, O. & Einhorn, S. (1994). Primary leukemia cells resistant to α-interferon in vitro are defective in the activation of the DNA-binding factor interferon-stimulated gene factor 3. Blood 84: 1942–1949.
Yamamoto, K., Quelle, F. W., Thierfelder, W. E., Kreider, B. L., Gilbert, D. J., Jenkins, N. A., Copeland, N. G., Silvennoinen, O. & Ihle, J. N. (1994). Stat4, a novel gamma interferon activation site-binding protein expressed in early myeloid differentiation. Mol Cell Biol 14: 4342–4349.
Zhang, P. & McLachlan, A. (1994). Differentiation-specific transcriptional regulation of the hepatitis B virus nucleocapsid gene in human hepatoma cell lines. Virology 202: 430–440.
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Hung, WC., Chuang, LY. Sodium butyrate enhances STAT 1 expression in PLC/PRF/5 hepatoma cells and augments their responsiveness to interferon-α. Br J Cancer 80, 705–710 (1999). https://doi.org/10.1038/sj.bjc.6690413
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bjc.6690413
Keywords
This article is cited by
-
Direct and indirect effects of IFN-α2b in malignancy treatment: not only an archer but also an arrow
Biomarker Research (2022)
-
Essential role of HCMV deubiquitinase in promoting oncogenesis by targeting anti-viral innate immune signaling pathways
Cell Death & Disease (2017)
-
Piperine induces apoptosis of lung cancer A549 cells via p53-dependent mitochondrial signaling pathway
Tumor Biology (2014)
-
Dual effects of sodium butyrate on hepatocellular carcinoma cells
Molecular Biology Reports (2012)
-
Differential effects of interferon alpha-2b and beta on the signaling pathways in human liver cancer cells
Journal of Gastroenterology (2005)