lncRNA MALAT1影响HIV-1感染与复制的分子机制研究

由于人类免疫缺陷病毒（HIV）能在CD4 T细胞中潜伏并形成病毒库，而高效抗逆转录疗法并不能有效清除病毒，因此了解调控HIV-1转录和潜伏分子机制对于艾滋病治疗尤为重要。长非编码RNA（lncRNA）的发现、鉴定及功能研究目前较集中在细胞发育、肿瘤发生等，而关于lncRNA与HIV-1功能及机制尚不清楚。前期利用RNA-Seq技术在HIV-1感染人CD4+T细胞中发现72个候选新lncRNA，qRT-PCR验证得到一个显著差异表达lncRNA MALAT1，且过表达MALAT1显著促进HIV-1感染。本项目拟进一步利用序列突变、流式细胞术、转录活性分析、RNA分析、蛋白印迹和RT-PCR等探讨MALAT1与HIV-1相互作用对病毒感染、病毒RNA稳定、复制等功能影响。本项目的完成将对探索lncRNA MALAT1功能、揭示新的HIV-1感染分子机制、为抗病毒治疗提供理论依据等具有重要意义。

As human immunodeficiency virus (HIV) could be persistent and latent infection in CD4 T cells and form viral reservior, highly active antiretroviral therapy didn’t effectively remove viruses. Therefore it is most important for AIDS treatment to understand the molecular mechanism on regulating transcription and latent infected by HIV-1. The detection and identification and function of new long non-coding RNA (lncRNA) is kept the focus on the cell development and tumorigenesis. It is largely unknown that the function and mechanism of lncRNAs acts on HIV-1 infection. Previously, we identified 72 novel lncRNAs in HIV-1 infection CD4+T cells by RNA-Seq. Interestingly, a novel lncRNA, metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) was expressed significantly increased in HIV-1 infected CD4+T cells compared to mock infected cells by qRT-PCR method. Moreover, we confirmed that over-expression of MALAT1 evidently resulted in increased HIV-1 replication by qRT-PCR and western blotting methods. In this study, the interaction and biding site/domain between HIV-1 and lncRNA MALAT1 will be further confirmed by the methods of northern blotting and sequencing mutational analysis. Then we will investigate how the interaction on HIV-1 and lncRNA MALAT1 effects viral infection and viral RNA stabilization and replication by the analysis of transcriptional activity, flow cytometry, western blotting and RT-PCR. It is great important significance that these findings will elucidate the new regulatory function of lncRNA MALAT1 and revel the molecular mechanism on HIV-1 infection, and will provide the helpful clue for anti-viral therapy.