MiR-10a*重组腺病毒促进B组3型柯萨奇病毒在心肌细胞的复制  

Ad-miRNA-10a Up-regulates Coxsackievirus B3 Replication in Cardiomyocytes

作　　者：佟雷[1] 胡盛林 屈允月 钟照华[1] Tong Lei;Hu Shenglin;Qu Yunyue;Zhong Zhaohua(Department ofMicrobiology,HarbinMedical University,Harbin,150080;Department of Education,The Open University of Harbin,Harbin,150001)

机构地区：[1]哈尔滨医科大学基础医学院,哈尔滨150080 [2]哈尔滨广播电视大学教学部,哈尔滨150001

出　　处：《基因组学与应用生物学》2020年第10期4845-4851,共7页Genomics and Applied Biology

基　　金：黑龙江省教育厅科学技术研究(面上)项目(12511176)资助。

摘　　要：为了明确miR-10a*在小鼠原代心肌细胞中对CVB3复制的作用,本研究在HEK293细胞中包装带有红色荧光(mCherry)并表达miR-10a*的重组腺病毒,将其感染HeLa细胞应用荧光显微镜观察荧光表达,RT-qPCR检测miR-10a*的表达;将Ad-miR-10a*感染HeLa细胞后,再感染CVB3,观察荧光表达,RTqPCR检测CVB3 RNA的表达;将Ad-miR-10a*感染小鼠原代心肌细胞再感染CVB3-eGFP,在不同时间点观察红色、绿色荧光表达;CVB3-eGFP感染小鼠原代心肌细胞,不同时间点观察绿色荧光表达,RT-qPCR检测miR-10a*的表达。本研究成功建立带有mCherry并表达miR-10a*的重组腺病毒(Ad-miR-10a*)并在HeLa细胞中观察到红荧光表达及检测到miR-10a*过表达。Ad-miR-10a*显著促进CVB3在HeLa中的复制;同样在小鼠原代心肌细胞中,Ad-miR-10a*可以促进CVB3的复制,并且Ad-miR-10a*感染细胞与CVB3感染细胞高度重合;CVB3-eGFP感染小鼠原代心肌细胞明显促进miR-10a*的表达。综上所述,Ad-miR-10a*可在HeLa、小鼠原代心肌细胞中过表达有生物学活性的miR-10a*;Ad-miR-10a*促进CVB3在原代心肌细胞中的复制,同时CVB3-eGFP促进miR-10a*在心肌细胞中的表达,CVB3可能通过此方式获得有利于自身复制和感染的环境,推测这可能是CVB3感染引起心肌疾病的重要机制之一。To investigate the effect of miR-10 a*on coxsackievirus group B3(CVB3)replication in cardiomyocytes.We construct and identify recombinant adenovirus(Ad-miRNA-10 a*)which express miR-10 a*and mCherry in Hela cells and cardiomyocytes.The mCherry expression was detected by fluorescence microscope.The expression of miRNAs were determined by RT-qPCR.Hela cells were infected with Ad-miRNA-10 a*and then infected with CVB3,the expression of CVB3 RNA was detected by RT-qPCR.The mice primary cardiomyocytes were infected with Ad-microRNA-10 a*and then infected with EGFP-CVB3.The expression of red and green fluorescence was observed at different time points by fluorescence microscope,and the expression of miRNA-10 a*was detected by RT-qPCR.We found that the recombinant adenovirus(Ad-miRNA-10 a*)with mCherry could be observed under fluorescence microscope and the overexpression of miRNA-10 a*was detected.Ad-mir-10 a*significantly promoted the replication of CVB3 in HeLa cells and mice primary cardiomyocytes.The expression of miRNA-10 a*were significantly promoted by EGFP-CVB3 infection in mice primary cardiomyocytes.Taken together,Ad-miR-10 a*can promote CVB3 replication in HeLa cells and mice primary cardiomyocytes.While CVB3 can promote the expression of miRNA-10 a*infection in cardiomyocytes.CVB3 may acquire an environment conducive to self-replication and infection by this way,which may be an important mechanism of CVB3 infection causing cardiomyopathy.

关 键 词：miR-10a* 重组腺病毒 B组3型柯萨奇病毒 心肌细胞 

分 类 号：R373.2[医药卫生—病原生物学]