EV71（Enterovirus 71）急性感染可引起手足口病及中枢神经系统损害，严重可导致死亡。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EV71病毒关系的功能、种类、数量及机制尚不清楚。我们前期工作利用RNA-Seq技术在EV71感染RD细胞中发现129个候选新lncRNA，qRT-PCR验证得到一个显著差异表达lncRNA STAM-AS1、且过表达STAM-AS1显著促进EV71感染。本项目拟进一步利用序列突变分析、Western blotting和RT-PCR等探索 STAM-AS1与EV71相互作用对病毒感染、复制等功能的影响。本项目完成将对探索长非编码RNA STAM-AS1功能、揭示新的EV71病毒感染分子机制以及为抗病毒治疗提供理论依据等都具有重要意义。

Enterovirus 71 (EV71) mostly affects children and causes hand, foot, and mouth disease with neurological complications. 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 classification and quantity and mechanism of lncRNAs acts on cells infected by EV71. Previously, we identified 129 novel lncRNAs in EV71 infected human malignant rhabdomyosarcoma (RD) cells by RNA-Seq. Interestingly, a novel lncRNA STAM-AS1 was expressed significantly increased in EV71 infected RD cells compare to mock infected cells by qRT-PCR method. Moreover, we confirmed that the overexpression of STAM-AS1 evidently resulted in increased EV71 replication by qRT-PCR and Western blotting method. In this study, their interaction and binding site/domain between EV71 and lncRNA STAM-AS1 will be further confirmed by the methods of Northern blotting and sequencing mutational analysis. Then we will investigate how the interaction of between EV71 and lncRNA STAM-AS1 effects viral infection and viral RNA stabilization and replication by the analysis of Western blotting and RT-PCR. It is great important significance that these finishing results will elucidate the new regulatory function of lncRNA STAM-AS1 and reveal EV71 infecting molecular mechanism, and will provide the helpful clue for anti-viral therapy.

EV71（Enterovirus 71）急性感染可引起手足口病及中枢神经系统损害，严重可导致死亡。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EV71病毒关系的功能、种类、数量及机制尚不清楚。本项目研究发现，EV71病毒感染能明显促进RD细胞中lncRNA STAM-AS1的表达，并且lncRNA STAM-AS1在EV71感染过程中发挥着重要的正向调控作用；过表达lncRNA STAM-AS1诱导正常RD细胞的自噬，并且lncRNA STAM-AS1能促进EV71对RD细胞自噬的诱导，这提示了lncRNA STAM-AS1可能通过对细胞自噬途径促进EV71的感染；lncRNA STAM-AS1 基因的494--1403bp区为重要功能决定区。本项目完成对探索长非编码RNA STAM-AS1功能、揭示新的EV71病毒感染分子机制以及为抗病毒治疗提供理论依据等都具有重要意义。