Malignant transformation of the cultured human normal biliary tract epithelial cells induced by hepatitis C virus core protein.

High level expression of hepatitis C virus core protein (HCV-C) was detected in hilar cholangiocarcinoma tissues in our previous studies. This protein played an important role in the process of cancer cell inversion and proliferation, by some direct and indirect effects on certain genes. Based on this observation, we investigated the effect of HCV-C on human normal biliary epithelial (hBE) cell transformation and tumor development. Plasmid pHCV-C encoding the gene of HCV core protein was constructed and transfected into hBE cells. The expression and the biological effect of HCV-C in HCV-C gene-modified hBE cells were determined in vitro and in vivo. The clone formation rates of hBE cells transfected with pHCV-C, pcdna3.1 and mock-transfected cells were 36, 2.5 and 1.5%, respectively. Tumor developed in 7 of 7 nude mice after incubated with pHCV-C transfected hBE cells, while no tumor appeared in mice injected with pcdna3.1- and mock-transfected hBE cells. To investigate the possible mechanism of malignant transformation, we further studied the telomerase activity and human telomerase reverse transcriptase (hTERT) expression in pHCV-C transfected hBE cells. The elevated expression of hTERT was confirmed by RT-PCR, immunocytochemistry (ICC) and Western blot analysis, which in turn elevated the telomerase activity, confirmed by TRAP-ELISA. These results indicated that HCV-C protein could participate in malignant transformation of human normal biliary epithelial cells and induce cholangiocarcinoma tumorigenesis, and the activation of telomerase was one of the possible mechanisms.

[1]  T. Patel,et al.  Interleukin-6 decreases senescence and increases telomerase activity in malignant human cholangiocytes. , 2006, Life sciences.

[2]  H. Chung,et al.  Alteration of hTERT full-length variant expression level showed different gene expression profiles and genomic copy number changes in breast cancer. , 2006, Oncology reports.

[3]  J. Wands,et al.  Hepatitis C virus core protein stimulates hepatocyte growth: Correlation with upregulation of wnt‐1 expression , 2005, Hepatology.

[4]  M. Colombo,et al.  Reconstitution of human telomerase reverse transcriptase expression rescues colorectal carcinoma cells from in vitro senescence: evidence against immortality as a constitutive trait of tumor cells. , 2005, Cancer research.

[5]  Ji-sheng Chen,et al.  Effect of hepatitis C virus core protein on modulation of cellular proliferation and apoptosis in hilar cholangiocarcinoma. , 2005, Hepatobiliary & pancreatic diseases international : HBPD INT.

[6]  A. Miyauchi,et al.  Telomerase activity in thyroid neoplasms evaluated by the expression of human telomerase reverse transcriptase (hTERT). , 2005, Anticancer research.

[7]  M. Manns,et al.  Telomeres and telomerase: A dual role in hepatocarcinogenesis , 2004, Hepatology.

[8]  S. Kubo,et al.  Hepatitis C virus infection as a likely etiology of intrahepatic cholangiocarcinoma , 2004, Cancer Science.

[9]  K. Tanimoto,et al.  Differential gene expressions during immortalization of normal human fibroblasts and endothelial cells transfected with human telomerase reverse transcriptase gene. , 2004, International journal of oncology.

[10]  G. Fontanini,et al.  Regulation of telomerase and its hTERT messenger in colorectal cancer. , 2004, Oncology reports.

[11]  Qi-bin Tang,et al.  Inhibitory effect of methylation inhibitor 5-aza-2-deoxycytidine on bile duct cancer cell line in vivo and in vitro. , 2004, Hepatobiliary & pancreatic diseases international : HBPD INT.

[12]  V. Lundblad.  Telomeres: Taking the measure , 2003, Nature.

[13]  S. Zou,et al.  Bile from a patient with anomalous pancreaticobiliary ductal union promotes the proliferation of human cholangiocarcinoma cells via COX-2 pathway. , 2003, World journal of gastroenterology.

[14]  N. Kelekis,et al.  Advanced intrahepatic cholangiocarcinoma in hepatitis C virus-related decompensated cirrhosis: case report and review of the literature. , 2003, European journal of gastroenterology & hepatology.

[15]  M. Kang,et al.  Elevated expression of hTERT is associated with dysplastic cell transformation during human oral carcinogenesis in situ. , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[16]  K. Hiroshima,et al.  Increased telomerase activity and elevated hTERT mRNA expression during multistage carcinogenesis of squamous cell carcinoma of the lung , 2001, Cancer.

[17]  M. Steffen,et al.  Motility factors identified in supernatants of human cholangiocarcinoma cell lines. , 2001, International journal of oncology.

[18]  S. Takeda,et al.  A case of the development of two hepatocellular carcinomas and a cholangiocarcinoma with cirrhosis after elimination of serum hepatitis C virus RNA with interferon therapy. , 2000, Hepato-gastroenterology.

[19]  K. Meyer,et al.  Hepatitis C virus core protein promotes immortalization of primary human hepatocytes. , 2000, Virology.

[20]  Y. Nakanuma,et al.  In situ nucleic acid detection of human telomerase in intrahepatic cholangiocarcinoma and its preneoplastic lesion , 1999, Hepatology.

[21]  K. Tsuchihara,et al.  Hepatitis C virus core protein regulates cell growth and signal transduction pathway transmitting growth stimuli. , 1999, Virology.

[22]  J. Southgate,et al.  Expression and cytokine regulation of immune recognition elements by normal human biliary epithelial and established liver cell lines in vitro. , 1998, Journal of hepatology.

[23]  Y. Konishi,et al.  Increased telomerase activity in intrahepatic cholangiocellular carcinomas induced by N-nitrosobis(2-oxopropyl)amine in hamsters. , 1998, Cancer letters.

[24]  Yoshiharu Matsuura,et al.  The core protein of hepatitis C virus induces hepatocellular carcinoma in transgenic mice , 1998, Nature Medicine.

[25]  S. Dash,et al.  HCV cDNA transfection to HepG2 cells , 1997, Journal of viral hepatitis.

[26]  K. Meyer,et al.  Hepatitis C virus core protein cooperates with ras and transforms primary rat embryo fibroblasts to tumorigenic phenotype , 1996, Journal of virology.

[27]  Robin C. Allshire,et al.  Telomere reduction in human colorectal carcinoma and with ageing , 1990, Nature.