Expression of nuclear factor kappa B in hepatitis C virus core gene transfect ed cholangiocarcinoma cells

OBJECTIVE: To establish an experimental model for exploring the role of hepatitis C virus (HCV) in the development of cholangiocarcinoma. METHODS: Recombinant plasmids of HCV-C gene were constructed by molecular cloning techniques and identified by PCR and restriction enzyme mapping.The plasmids were then transfected into QBC939 cells (a cholangiocarcinoma cell line) by Lipofection. After selection with G418, resistant colonies were obtained and analyzed by immunocytochemistry and Western blotting. The morphology was observed by trans mission electron microscopy (TEM). The expression of NF-(k)B was detected by immunocytochemistry. RESULTS: Recombinant plasmid was shown by PCR and restriction enzyme mapping to carry the target gene. Moreover, it could efficiently express HCV-C protein in QBC939 cells. HCV-like particles were found in the cytoplasm by TEM, which were spherical with a diameter of 50-80 nm and possessed an outer membrane. Moreover, NF-(k)B activation could be shown in HCV core-transfected cells. CONCLUSION: Expression of the HCV-C gene in cholangiocarcinoma cells was achieved. Transfected tumor cells (QBC939-HCVc) could be used as a model to study the effect of HCV on the development of cholangiocarcinoma.