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目的 以乙型肝炎病毒(HBV)核心区为靶位,构建表达小干扰RNA(siRNA)的质粒载体pSilencer3.1-Hlhygro,体外观察siRNA抗HBV的效果。 方法 以HepG2 2.2.15细胞为靶细胞,利用脂质体Metafectene与表达siRNA的质粒载体pSilencer3.1-Hlhygro共转染,用定量聚合酶链反应检测细胞上清液中DNA,用逆转录聚合酶链反应检测HBV C-mRNA。 结果 成功构建了表达siRNA的转录质粒载体,两条siRNA均可抑制HBV的复制,而且与siRNA浓度成正相关。 结论 靶向HBV核心区的siRNA能抑制HBV的复制。
Objective To construct a plasmid vector pSilencer3.1-Hlhygro expressing small interfering RNA (siRNA) targeting the core of Hepatitis B virus (HBV), and to observe the effect of siRNA against HBV in vitro. Methods HepG2 2.2.15 cells were used as target cells and cotransfected with pSilencer3.1-Hlhygro, a plasmid vector expressing lipofectamine. The DNA in supernatant was detected by quantitative polymerase chain reaction (RT-PCR) Chain reaction detection of HBV C-mRNA. Results The siRNA plasmid vector was successfully constructed. Both siRNAs could inhibit the replication of HBV and was positively correlated with the siRNA concentration. Conclusion siRNA targeting HBV core can inhibit the replication of HBV.