宿主miRNA-2127靶向p53促进禽流感病毒H9N2亚型体外复制的分子机制  

Molecular Mechanism of Host miRNA-2127 Targeting p53 Promoting H9N2 Subtype of Avian Influenza Virus Replication in vitro

作　　者：张玉霞[1] 袁小远[1] 杨金兴[1] 孟凯[1] Zhang Yuxia;Yuan Xiaoyuan;Yang Jinxing;Meng Kai(Poultry Institute,Shandong Academy of Agricultural Sciences,Jinan 250100,China)

机构地区：[1]山东省农业科学院家禽研究所

出　　处：《山东农业科学》2019年第12期91-95,共5页Shandong Agricultural Sciences

基　　金：山东省自然科学基金项目(ZR2017BC039);山东省重点研发计划项目(2018GSF121012);山东省重点实验室专项建设计划项目(SDKL201810);山东省农业科学院家禽研究所学科团队建设项目(CXGC2018E11)

摘　　要：为研究鸡体内miRNA-2127对低致病性禽流感病毒(avian influenza virus,AIV)H9N2复制的影响,本研究将化学合成的gga-miR-2127模拟物和抑制剂分别转染DF-1细胞,接种H9N2亚型病毒,检测细胞中病毒含量变化,发现gga-miR-2127能够显著促进病毒的复制。为探索其中的分子机制,用生物信息学软件预测gga-miR-2127的调控位点位于p53基因mRNA的3′UTR区,并用双荧光素酶报告系统对其靶向关系进行了验证。荧光定量RT-PCR法和Western blot试验表明,gga-miR-2127过表达时p53的mRNA水平不变而p53蛋白表达水平降低,细胞天然免疫机能下降。据此推测gga-miR-2127促进H9N2复制的分子作用机理是在p53基因的mRNA转录后,通过gga-miR-2127与其mRNA的3′UTR区结合,抑制p53蛋白的翻译从而抑制抗病毒作用和细胞的天然免疫机能。To study the effect of miRNA-2127 on the replication of low pathogenic AIV(avian influenza virus)subtype H9N2 in vitro,chemically synthesized gga-miR-2127 mimic and inhibitor were transfected into DF-1 cells inoculated with H9N2 subtype,and the changes of virus content in cells were detected.It was found that gga-miR-2127 could significantly promote the replication of AIV virus.In order to explore the molecular mechanism,the regulatory site of gga-miR-2127 was predicted as 3′UTR region of mRNA of p 53 gene by bioinformatics software,and the targeting relationship was validated by double luciferase reporting system.Fluorescence quantitative RT-PCR and Western blot assay showed that the mRNA of p53 gene remained unchanged while the expression of p53 protein decreased under the condition of overexpressing gga-miR-2127,and the innate immune function of cells decreased.It was speculated that the molecular mechanism of gga-miR-2127 promoting H9N2 replication was through inhibiting the translation of p53 protein by binding with the 3′UTR region of p53 gene after transcription,so the antiviral effect and the realization of cell innate immune function were inhibited.

关 键 词：H9N2亚型禽流感病毒 gga-miR-2127 分子机制 

分 类 号：S852.65[农业科学—基础兽医学]