RNA干扰抑制HBV e抗原基因表达的实验研究

Inhibitory effect of RNAi on HBV preC/C gene expression

目的针对HBV前C/C基因的siRNA表达载体pSiHBV/C、含HBV前C/C基因(e抗原基因)及绿色荧光蛋白基因的融合表达载体pEGFP-HBVC(1或4),共转染真核细胞,观察RNA干扰对HBVe抗原基因表达的抑制作用。方法脂质体共转染pSiHBV/C和pEGFP-HBVC真核细胞,通过观察荧光强度,及利用蛋白免疫印迹进行半定量分析来检测RNA干扰作用对HBVe抗原基因表达的抑制效果。结果针对HBV前C/C基因的siRNA表达载体pSiHBV/C与pEGFP-HBVC共转染真核细胞后,可明显抑制融合蛋白的表达。结论在细胞水平上,针对HBV前C/C基因的siRNA表达载体pSiHBV/C产生的siRNA可特异的抑制HBVe抗原基因的表达。

[Objectives] To evaluate the inhibitory effect of RNAi on HBV preC/C gene. [Methods] The RNAi plasmid pSiHBV/C（1 or 4） and the recombinent plasmid pEGFP-HBVC were cotransfected into HEK293 cells tho assess the inhibitory effect of RNAi on HBV preC/C gene expression by means of ELISA and western blotting. [Resuits] The expression siRNA vector pSiHBV/C targeting the core gene of HBV inhibited the expression of double function fusion protein efficiently and specifically. [Conclusion] At the cell level, inhibition of HBeAg gene expression by RNAi plasmid pSiHBV/C provides the foundation for further study about RNAi targeting HBeAg.