反义RNA抑制乙肝病毒复制的临床分子生物学研究

lnhibition of hepatitis B virus by retroviral vectors expressing antisense RNA

目的 研究表达反义RNA的逆转录病毒载体体外转基因抗乙肝病毒 (HBV)的作用 ,为临床治疗乙型肝炎探索一条新途径。方法 合成互补于HBVX区 (140 0 - 1430 )的X片段 ,互补于HBVP区 (2 375 - 2 40 5 )的P片段 ,分别构建表达正、反义X或P的重组逆转录病毒载体质粒PLXSN +Xpos/Ppos,PLXSN +X ,PLXSN +P ,转染PA317细胞 ,用假病毒颗粒感染2 2 15细胞后第 8天 ,提取细胞内DNA和RNA ,用荧光PCR(FQ -PCR)定量方法检测 2 2 15细胞的DNA和RNA。结果 反义RNA对HBVS、C、P三个开放读码区的DNA及RNA抑制作用大。结论 细胞内表达HBV反义RNA是有明显抗乙肝病毒复制和表达的作用 。

Objective Research on the effects of antisense RNA against HBV infection.Methods Retroviral vector expressing antisense RNA complementary to HBVX(1400-1430),P(2375-2405)was used to transduce 2.2.15 cell.We use FQ-PCR methods to test celluar HBV DNA and HBV RNA.Results The results showed that the celluar DNA and RNA in S、C、P ORFs were greatly inhibited.Conclusions The results demonstrated that retroviral vectors containing shorter antisense fragment can inhibit HBV replication and expression.Therefore the antisense RNA may be potentially useful for anti-HBV gene therapy. [