RNA干扰抑制麻疹病毒体外复制的实验研究

为了研究短发夹RNA(shRNA)介导的RNA干扰对麻疹病毒体外复制的抑制作用,构建靶向与麻疹病毒复制密切相关的宿主细胞基因Rab9 GTPase基因特异性shRNA表达载体,分别转染Vero-E6和B95a细胞后感染麻疹病毒Edmonston株和野生株。逆转录聚合酶链反应(RT-PCR)和免疫印迹技术(Western-blot)检测转染细胞内Rab9 GTPase基因表达水平;标准蚀斑试验测定麻疹病毒滴度。结果显示转染细胞内Rab9 GTPase mRNA和蛋白质的表达水平同对照组相比明显降低,标准蚀斑试验显示麻疹病毒的复制受到显著抑制,抑制率达到90%以上。结果表明载体介导的shRNAs能通过特异性下调Rab9 GTPase基因表达抑制麻疹病毒体外复制,Rab9 GTPase可能成为治疗麻疹病毒感染的RNA干扰靶。

Inhibition of Measles Virus Replication by RNA Interference in vitro

In order to investigate the inhibitory effect of RNA interference via vector-based specific short hairpin RNA （shRNA） on measles virus replication in vitro, two shRNA expression vectors targeting Rab9 GTPase gene of host cell and closely related to measles virus replication were constructed and transfected into Vero-E6 and B95a cells, then infected the cells by measles virus Edmonston strain and wild-type strain respectively. The expression level of Rab9 GTPase mRNA in the transfected cells was assayed by RT-PCR and Western-blot. The titers of measles virus were determined by standard plaque assays. The results showed that shRNA targeting Rab9 GTPase gene could specially inhibited the expression level of Rab9 GTPase mRNA and protein in culture cells; Measles virus replication was obviously suppressed by two shRNAs targeting Rab9 GTPase gene, with over 90% of inhibition rate. These results demonstrated that vector-based shRNA could suppress measles virus replication in vitro through specially down-regulating the expression of Rab9 GTPase and Rab9 GTPase might become a good RNA interference target against measles virus infection.