Abstract:Objective:To investigate the effect of hepatitis c virus NS3 gene on the expression of phosphorylate tyrosine proteins in human hepatocyte.
Methods :HCV NS3 plasmids and pRcCMV plasmids were stably transfected into QSG7701 cells, which were named as pRcHCNS3/QSG cells and pRcCMV/QSG cells respectively. Western Blot and immunocytochemistry was used to detect the expression of phospho-p44/42, phospho-p38 and phospho-SAPK/JNK in the pRcHCNS3/QSG cells, pRcCMV/QSG cells and QSG7701 cells, respectively. Immunoprecipitation analysis was employed to investigate the effect of HCV NS3 on the phosphotyrosine protein expression.
Results:The expressions of phospho-p44/42, phospho-p38 and phospho-SAPK/JNK were found in pRcHCNS3/QSG cells, pRcCMV/QSG cells and QSG7701 cells. The positive signals of phospho-p44/42, phospho-p38 and phospho-SAPK/JNK protein were localized in the nucleus of cells. The signals intensity of the pRcHCNS3/QSG cells was stronger than the other the two cells. The expression of phosphotyrosine protein was also up-regulated in pRcHCNS3/QSG cells.
Conclusions:HCV NS3 may affect cell signaling pathway through activating tyrosine kinase and followed with activation and abnormal phosphorylation of several cytoplasmic signaling molecules such as MAPKs, and induce unlimited hepatocyte transformation and tumor development.