抗乙型肝炎病毒(HBV)S区的siRNA对HBV-DNA的抑制作用

体外观察s1RNA对HepG2 2.2.15细胞中乙型肝炎病毒(HBV)HBV-DNA分泌的影响。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 ZYl、pSilencer ZY2和pSilencer ZY3,测序及酶切鉴定后用脂质体介导重组质粒转梁HepG2 2.2.15细胞,用酶免分析法对HepG2 2.2.15 细胞上清中HBV-DNA进行定量检测。结果成功构建了针对HBV S区的siRNA的表达载体,三条s1RNA 均可不同程度地抑制HepG2 2.2.15细胞上清中HBV-DNA,转染72 h抑制效果最好,分别为62%、31%、63%。说明针对HBV S区的siRNA能明显抑制HBV-DNA。

Inhibition of HBV-DNA by siRNA Targeting HBV S Antigen

To observe the inhibitory effect of siRNA on HBV DNA in HepG2 2. 2. 15 cells, three siRNAs targeting the S-RNA of HBV were synthesized and cloned into pSilencer ZY1, pSilencer ZY2, pSilencer ZY3 and pSilencer HK. Transfection into HepG2 2. 2. 15 cells with the new recombinant vectors was carried out by Metafectene. The HBV DNA in the supernatant of HepG2 2. 2. 15 cells was assayed by Fluorescence Quantitative PCR. The construction of the recombinant expression vectors was successfully conformed by the results of enzyme digestion, electrophoresis and gene sequencing. The three siRNAs inhibited HBV DNA in the supernatant of HepG2 2. 2. 15 cells by 62%, 31% and 63% respectively at 72 h after transfection. It is conclused that the three siRNAs mentioned above targeting the S-mRNA of HBV could effectively inhibit HBV DNA.