Abstract
The present study was designed to research on RNA interference hepatitis B virus x gene approach to hepatocellular carcinoma (HCC) therapy. Previously, we constructed and identified shRNA eukaryotic expression vectors (pshRNA-X220) specific to HBx gene, pshRNA-MOCK (control); and established HCC cell lines with stable expression shRNA eukaryotic vector targeting HBx gene—21543 cell lines (MHCC97-H of expressing shRNA against HBx), HK3 cell lines (MHCC97-H by transfected with pshRNA-MOCK). We examined the expression of HBx gene after RNA interference by semi-quantitative RT-PCR and assessed the effect of HBx knocked down on cell growth by proliferation assay using kit-8 (CCK8). As well as, we analyzed cell cycle distribution by flowcytometry and examined cell apoptosis using TUNEL assay. The HBx mRNA expression level is reduced, and cells growth was significantly stopped in 21543 cell lines. Cells with HBx knockdown were more sensitive to 5-fluorouracil/cisplatin. RNA interfering HBx induced an obvious time and dose-dependent inhibitory in comparison with the control cells. Meanwhile, RNA interferenced targeting HBx, in combination with chemotherapy can effectively induce apoptosis in hepatocellular carcinoma cells and restricts cell proliferation.
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Wang WL, London WT, Feitelson MA. Hepatitis B × antigen in hepatitis B virus carrier patients with liver cancer. Cancer Res. 1991;51(18):4971–7.
Feitelson MA, Duan LX. Hepatitis B virus X antigen in the pathogenesis of chronic infections and the development of hepatocellular carcinoma. Am J Pathol. 1997;150(4):1141–57.
Nowak AK, Chow PK, Findlay M. Systemic therapy for advanced hepatocellular carcinoma: a review. Eur J Cancer. 2004;40(10):1474–84.
Wang WL, et al. Identification and construction of shRNA for targeted HBx gene integrated to MHCC-97H cell line. J China medical univ. 2008;37(4):445–7.
Xing-e HE, et al. Establishment of human hepatocellular carcinoma cell line expressing HBx shRNA. Zhonghua Gan Zang Bing Za Zhi. 2009;17(4):310–2.
Michielsen PP, Francque SM, van Dongen JL. Viral hepatitis and hepatocellular carcinoma. World J Surg Oncol. 2005;3:27–32.
Farazi PA, DePinho RA. Hepatocellular carcinoma pathogenesis: from genes to environment. Nat Rev Cancer. 2006;6(9):674–87.
Haviv I, Shamay M, Doitsh G, Shaul Y. Hepatitis B virus pX targets TFIIB in transcription coactivation. Mol Cell Biol. 1998;18(3):1562–9.
Milich DR, et al. Is a function of the secreted hepatitis B e antigen to induce immunologic tolerance in utero ? Proc Natl Acad Sci U S A. 1990;87(17):6599–603.
Wei Y, et al. Identification of beta-1, 4-galactosyltransferase I as a target gene of HBx-induced cell cycle progression of hepatoma cell. J Hepatol. 2008;49(6):1029–37.
Benn J, Schneider RJ. Hepatitis B virus HBx protein deregulates cell cycle checkpoint conrols. Proc Natl Acad SCI U S A. 1995;92(24):11215–9.
DeVincenzo JP. RNA interference strategies as therapy for respiratory viral infections. Pediatr Infect Dis J. 2008;27(10):S118–22.
Fire A, et al. Potent and specific genetic interference by double stranded RNA in Caenorhabditis elegans. Nature. 1998;391(6669):806–11.
Giladi H, et al. Small interfering RNA inhibits hepatitis B virus replication in mice. Mol Ther. 2003;8(5):769–76.
Romano PR, McCallus DE, Pachuk CJ. RNA interference-mediated prevention and therapy for hepatocellular carcinoma. Oncogene. 2006;25(27):3857–65.
Cheng AS, et al. RNA interference targeting HBx suppresses tumor growth and enhances cisplatin chemosensitivity in human hepatocellular carcinoma. Cancer Lett. 2007;253(1):43–52.
Schulze-Bergkamen H, et al. Suppression of mcl-1 via RNA interference sensitizes human hepatocellular carcinoma cells towards apoptosis induction. BMC Cancer. 2006;6:232–40.
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He, Y., Sun, Hq., He, Xe. et al. Knockdown of HBx by RNAi inhibits proliferation and enhances chemotherapy-induced apoptosis in hepatocellular carcinoma cells. Med Oncol 27, 1227–1233 (2010). https://doi.org/10.1007/s12032-009-9363-0
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DOI: https://doi.org/10.1007/s12032-009-9363-0