目的研究La蛋白与乙型肝炎病毒(HBV)mRNA稳定性和蛋白质表达之间的相关性。方法设计并合成La蛋白mRNA特异性小干扰RNA(siRNA),转染HepG2.2.15细胞,继续培养3d后分别裂解转染和未转染siRNA的细胞l抽提蛋白质,Westernblot检测La蛋白含量变化;于转染后的第1、2、3、5、7、9天分别收集细胞培养上清液,实时荧光定量聚合酶链反应技术检测上清液中HBVDNA的变化情况,电化学发光分析技术检测乙型肝炎表面抗原,乙型肝炎e抗原含量变化情况。结果La蛋白在转染siRNA的HepG2.2.15细胞中表达明显低于未转染siRNA的细胞;转染siRNA的细胞分泌到上清液中乙型肝炎表面抗原、乙型肝炎e抗原和HBVDNA的含量也明显低于未转染siRNA的细胞。结论在HepG2.2.15细胞内,La蛋白与HBVmRNA稳定性和蛋白质表达具有功能相关性。
Objective To study the relativity between La protein and the stability of HBV mRNA and the expression of HBV protein. Methods Four specific siRNAs were obtained by transcription in vitro. After transfection with the siRNAs into HepG2.2.15 cells for 3 days, the inhibitive effects of La protein were analyzed by Western blot; the content changes of HBsAg, HBeAg and HBV-DNA were detected by ECL and RT-PCR. Results In comparison to normal cells, La protein was less in the cells. There was less La protein in the cells transinfected with siRNAs. HBsAg, the HBeAg and HBV-DNA secreted by the cells transfected with sLRNA were also less than that in the normal cells. Conclusion There is a correlation between La protein and HBV mRNA and the expression of HBV protein.