Expression of nuclear factor kappa B in hepatitis C virus core gene transfected 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.


CONCLUSIONS
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.
Liu
Xiaofang
X
Department of General Surgery of Tongji Hospital, Wuhan 430030, China. Liu634@263.net
Zou
Shengquan
S
Qiu
Fazu
F
D001650:Bile Duct Neoplasms / Q000737:chemistry / Q000209:etiology / Q000821:virology*; D001653:Bile Ducts, Intrahepatic; D018281:Cholangiocarcinoma / Q000737:chemistry / Q000209:etiology / Q000821:virology*; D006801:Humans; D016328:NF-kappa B / Q000032:analysis*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D014162:Transfection; D014407:Tumor Cells, Cultured; D014758:Viral Core Proteins / Q000235:genetics / Q000502:physiology*
D016328:NF-kappa B; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus