Construction of HCV-core gene vector and its expression in cholangiocarcinoma.
OBJECTIVE
To establish an experimental model for exploring the role of hepatitis C virus (HCV) in the development of cholangiocarcinoma.


METHODS
Recombinant plasmid of HCV-core gene was constructed with molecular cloning technique and transfected into QBC939 cells with lipofection.After it was selected with G418, resistant colonies were obtained. The colonies were analysed by immunocytochemistry and Western blotting. The morphology was observed under transmission electron microscope(TEM) and microscope.


RESULTS
The recombinant plasmid was proved to carry the target gene by PCR and restriction enzymed mapping. Moreover, it could express HCV-C protein efficiently in QBC939 cells. The HCV-like particles were found in the cytoplasm by electron microscope, which were spherical with a diameter of 50 nm-80 nm possessing outer membrane. The transfected cells had lower differentiation and higher malignant degree under microscope.


CONCLUSIONS
Because HCV-core gene could express steadily in cholangiocarcinoma cells,the transfected tumor cells(QBC939-HCVC) could be used to study the effect of HCV in the development of cholangiocarcinoma.
Liu
Xiao-Fang
XF
Department of General Surgery of Tongji Hospital, 1095 Jiefang Road, Wuhan 430030, Hubei Province,China. liu634@263.ne
Zou
Sheng-Quan
SQ
Qiu
Fa-Zu
FZ
D001650:Bile Duct Neoplasms; D001653:Bile Ducts, Intrahepatic; D018281:Cholangiocarcinoma; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D008854:Microscopy, Electron; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000648:ultrastructure / Q000821:virology; D014758:Viral Core Proteins / Q000235:genetics
D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus