Summary: | 碩士 === 慈濟大學 === 分子生物及細胞生物研究所 === 90 === Infection with hepatitis C virus (HCV) could cause chronic hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC). How HCV induces HCC is largely unknown. In transgenic mice model, HCV core protein alone could induce HCC. The purpose of this thesis is to study the effect of HCV core protein on a hepatoblastoma cell line, HuH7 cells. To obtain this goal, we have constructed HCV core protein expression plasmid, and selected stable transformants in HuH7 cells using G418. These stable transformants were used to study the effect of HCV core protein on HuH7 cells. The expression of tumor suppressor p53 was similar between parental HuH7 cells and stable transformants with HCV core protein (HuH7/CC-3). In normal culture condition, the growth rate of HuH7/CC-3 stable transformant was also similar to that of HuH7 cells. On the other hand, when cultured in serum free condition (DMEM only), survival cells from HuH7/CC-3 stable transformant were more than those from HuH7 cells. Apoptosis induced by serum starvation in HuH7 and HuH7/CC-3 cells was demonstrated by Hoechst staining assay.Therefore, HCV core protein could inhibit serum starvation-induced apoptosis. It has been reported that HCV core protein could activate NF-κB. It also has been demonstrated that activation of NF-κB could inhibit apoptosis. To prove that HCV core protein could inhibit apoptosis induced by serum starvation through NF-κB, NF-κB inhibitor (curcumin) was added in the serum-free medium. Survival cells of HuH7/CC-3 were reduced when treated with NF-κB inhibitor. From these results, we conclud that HCV core protein inhibits apoptosis induced by serum-starvation through NF-κB activation.
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