HepG2.2.15细胞中siRNA与拉米夫定抗乙型肝炎病毒复制作用的比较

Comparison of the inhibitory effect on HBV replication by siRNA and lamivudine in HepG2.2.15 cells

目的:对HepG2.2.15细胞中siRNA与拉米夫定的抗乙型肝炎病毒(HBV)作用进行比较。方法:构建HBV siRNA的表达载体并转染入HepG2.2.15细胞。分别于48、72、96h收获细胞,与拉米夫定治疗组比较抗HBV作用。用ELISA方法检测HBsAg浓度;HBV DNA水平用实时定量PCR测定;用逆转录PCR检测HBV mRNA水平。结果:拉米夫定能明显降低HBV DNA水平,对HBsAg和mRNA抑制率较低;siRNA能全面降低HBsAg、HBV DNA和mRNA水平(P<0.05)。结论:HepG2.2.15细胞中,siRNA与拉米夫定的抗病毒作用完全不同,siRNA抗HBV作用明显优于拉米夫定。

AIM：To compare the inhibitory effect of inhibition of siRNA and lamivudine on hepatitis B virus replication and expression in HepG2.2.15 cells. METHODS： Recombinant plasmid psil-HBV was constructed and transfected into HepG2.2.15 cells. The transfected cells were harvested at 48,72 and 96 h. The concentration of HBeAg and HBsAg was determined using ELISA method. HBV DNA replication was examined by real time PCR, and the level of HBV mRNA was measured by reserved transcribed PCR.RESULTS：siRNA effectively inhibited the expressing and replication of HBV at the levels of HBsAg, HBV DNA and mRNA（P0.05）. Lamivudine had only inhibitory effect on viral DNA synthesis. CONCLUSION： The anti-HBV effect of HepG2.2.15 cells treated by siRNA is totally differently and more effective from that seen with lamivudine in HepG2.2.15 cells.