针对HBVS区的siRNA的抗病毒活性

目的: 体外观察siRNA对HepG2 2.2.15细胞中乙型肝炎病毒(HBV)S-mRNA及HBsAg,HBeAg产生的影响. 方法: 设计并合成针对HBV S区的三条siRNA,构建含上述siRNA的表达载体pSilencer ZY1,pSilencer ZY2和pSilencer ZY3,pSilencer HK2测序及酶切鉴定后用脂质体介导重组质粒转染HepG2 2.2.15细胞,用酶免分析法对HepG2 2.2.15细胞上清中HBsAg,HBeAg进行检测,用逆转录聚合酶链反应检测HBV S-mRNA.结果: 成功构建了针对HBV S区的siRNA的表达载体,三条siRNA均可程度不同地抑制HepG2 2.2.15细胞上清中HBsAg,HBeAg的分泌,72 h抑制率达高峰,对HBsAg的抑制率分别为81%,29%及78%,对HBeAg的抑制率分别为35%,3%及49%,并有抑制HBV S-mRNA的作用.结论: 针对HBV S区的siRNA能明显抑制HBV的复制.

Anti-HBV effect of siRNA expression plasmid targeting the S antigen of HBV

AIM: To identify the siRNA interference ability for the replication of HBV. METHODS: Based on the sequence of HBV in GenBank, siRNA targeting the S-RNA of HBV were synthesized and cloned into pSilencer ZY1, pSilencer ZY2, pSilencer ZY3 and pSilencer HK. Transfected into HepG2 2.2. 15 cells with the new recombinant vectors was carried out by Metafectene . The expression of HBsAg and HBeAg was assayed by MEIA and HBV S-mRNA was detected by RT-PCR. RESULTS: The construction of the recombinant expression vectors was successfully conformed by the results of enzyme digestion, electrophoresis and gene sequencing. The three siRNAs could suppress HBV S-mRNA and the expression of HBsAg and HBeAg in the supernatant of HepG2 2.2.15 cells. The inhibitory rates of HBsAg were 81%, 29% and 78%, and that of HBeAg were 35%, 3% and 49% respectively. CONCLUSION: The three siRNA mentioned above targeting the S-mRNA of HBV could effectively inhibit HBV replication.