目的:探讨过表达热休克蛋白70(HSP70)对乙型肝炎病毒(HBV)复制的影响。方法:运用RTq PCR检测HBV复制对HSP70表达的影响及过表达HSP70对HBV 3.5 kb m RNA的影响;运用Western blot法验证HSP70过表达效果及过表达HSP70对HBV核心蛋白的影响;RT-q PCR和Southern blot法分析过表达HSP70对HBV复制中间体的影响;运用双萤光素酶报告系统检测过表达HSP70对HBV启动子活性的影响。结果:HBV复制显著抑制HSP70的m RNA水平。过表达HSP70抑制HBV复制中间体、3.5 kb m RNA以及核心蛋白的表达,同时抑制HBV核心启动子的活性。结论:HBV复制抑制HSP70的表达。过表达HSP70抑制HBV的复制。本研究结果表明,HSP70通过抑制核心启动子的活性抑制HBV的复制。
AIM: To investigate the role of heat shock protein 70(HSP70)in hepatitis B virus(HBV) replication. METHODS: The effect of HBV replication on the expression of HSP70 was analyzed by RT-q PCR. The overexpression efficiency of HSP70 was confirmed by Western blot. The effect of HSP70 overexpression on HBV DNA replicative intermediates was analyzed by RT-q PCR and Southern blot. The effects of HSP70 overexpression on the expression level of HBV 3. 5 kb m RNA and HBV core protein were measured by RT-q PCR and Western blot,respectively. The Effect of HSP70 overexpression on HBV promoter activity was detected by dual luciferase reporter system. RESULTS: The m RNA levels of HSP70 were inhibited by HBV replication. Overexpression of HSP70 repressed the expression of HBV DNA replicative intermediates,3. 5 kb m RNA and core protein,as well as HBV core promoter activity. CONCLUSION: HBV replication inhibits the expression of HSP70. Overexpression of HSP70 represses HBV replication. These data suggest that HSP70 repressed HBV replication by inhibiting HBV core promoter activity.