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AIM:To establish an experimental model for exploring therole of hepatitis C vivus(HCV)in the development ofchofangiocarcinoma.METHODS:Recombinant plasmid of HCV-core gene wasconstructed with molecular cloning technique andtransfected into QBC939 calls with lipofection.After it wasselected with G418,resistant colonies were obtained.Thecolonies ware analysed by immunocytochemistry andWestern blotting.The morphology was observed undertransmission electron microscope(TEM)and microscope.RESULTS:The recombinant plasmid was proved to carry thetarget gene by PCR and restriction enzymed mapping.Moreover,it could express HCV-C protein efficiently inQBC939 cells.The HCV-like particles were found in thecytoplasm by EM,which ware spherical with a diameter of50nm-80nm possessing outer membrane.The transfectedcells had lower differentiation and higher malignant degreeunder microscope.CONCLUSION:Because HCV-core gene could expresssteadily in cholangiocarcinoma calls,the transfected tumorcells(QBC939-HCVC)could be used to study the effect ofHCV in the development of cholangiocarcinoma.
AIM: To establish an experimental model for exploring the blood of hepatitis C vivus (HCV) in the development of chofangiocarcinoma. METHODS: Recombinant plasmid of HCV-core gene wasconstructed with molecular cloning technique and transferred into QBC939 calls with lipofection. After it wasselected with G418, resistant colonies were obtained. The colony ware was analyzed by immunocytochemistry and Western blotting. The morphology was observed in electron microscope (TEM) and microscope. RESULTS: The recombinant plasmid was proved to carry the target gene by PCR and restriction enzym mapping. Moreover, it could express express HCV- C protein efficiently in QBC939 cells. The HCV-like particles were found in the cytoplasm by EM, which ware spherical with a diameter of 50nm-80nm possessing outer membrane. The transfected cells had lower differentiation and higher malignant degreeunderunder microscope. CONCLUSION: Because HCV-core gene could expresssteadily in cholangiocarcinoma calls, the transfected tumor cells (QBC939-H CVC) could be used to study the effect of HCV in the development of cholangiocarcinoma.