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Objective:To investigate the effect of hepatitis C virus non-structural protein 4B(HCV NS4B) on c-Myc, P53, ras gene expression and apoptosis in hepatic cells and study the possible role that NS4B played in the carcinogenesis of hepatoma. Methods: The recombinant plasmid(PCXN2-NS4B, PCXN2-P53) and the empty vector were transfected or co-transfected into Chang liver cells with liposome. Screening was performed with G418. Plasmid mRNA was detected by RT-PCR. The protein expressions of c-Myc and ras genes were analyzed by immunocytochemistry. The expressions of wild-type P53(wtp53) gene were detected by in situ hybridization. TUNEL(flow cytometry) was used for assessing the rate of apoptosis. Results:No expression of c-Myc gene was found in PCXN2 group. The expression of c-Myc gene in NS4B group was 21.3% ± 1.2%. The expression of ras gene in PCXN2 group was lower than that in NS4B group. Compared with PCXN2 group, the expression of P53 mRNA was not promoted or inhibited in NS4B group. But the expression of P53 mRNA in NS4B-P53 group was lower than that in P53 group. In PCXN2, NS4B, P53 and NS4B-P53 group, the rates of apoptosis were 17.02% ± 1.24%, 11.94% ± 2.24%, 25.84% ± 3.49% and 18.34% ± 1.55% respectively. Conclusion:HCV NS4B induces the expression of c-Myc and ras gene. HCV NS4B may play a role in the inhibition of cell death through P53-dependent manner. Results from this study suggested that HCV NS4B might contribute to the viral carcinogenesis.
Objective: To investigate the effect of hepatitis C virus non-structural protein 4B (HCV NS4B) on c-Myc, P53, ras gene expression and apoptosis in hepatic cells and study the possible role that NS4B played in the carcinogenesis of hepatoma. Methods: The recombinant plasmid (PCXN2-NS4B, PCXN2-P53) and the empty vector were transfected or co-transfected into Chang liver cells with liposome. Screening was performed with G418. Plasmid mRNA was detected by RT-PCR. The protein expressions of c- The expressions of wild-type P53 (wtp53) gene were detected by in situ hybridization. TUNEL (flow cytometry) was used for assessing the rate of apoptosis. Results: No expression of c-Myc gene was found in PCXN2 group. The expression of c-Myc gene in NS4B group was 21.3% ± 1.2%. The expression of ras gene in PCXN2 group was lower than that in NS4B group. not promoted or inhibited in NS4B group. B The rates of apoptosis were 17.02% ± 1.24%, 11.94% ± 2.24%, 25.84% in the NS4B-P53 group was lower than that in P53 group. In PCXN2, NS4B, P53 and NS4B- ± 3.49% and 18.34% ± 1.55% respectively. Conclusion: HCV NS4B induces the expression of c-Myc and ras gene. HCV NS4B may play a role in the inhibition of cell death through P53-dependent manner. Results from this study suggested that HCV NS4B may contribute to the viral carcinogenesis.