柯萨奇病毒B3型感染引起HeLa细胞内源性小干扰RNA的变化

姚海兰1; 宋娟2; 王欣玲2; 王瑞芳2; 宋芹芹2; 史冰田2; 韩俊2

doi:10.3969/j.issn.1673-6184.2020.02.005

微生物与感染 >

2020 , Vol. 15 >Issue 2: 98 - 103

DOI: https://doi.org/10.3969/j.issn.1673-6184.2020.02.005

论著

柯萨奇病毒B3型感染引起HeLa细胞内源性小干扰RNA的变化

姚海兰1 ,
宋娟2 ,
王欣玲2 ,
王瑞芳2 ,
宋芹芹2 ,
史冰田2 ,
韩俊2

展开

1. 首都儿科研究所生物化学与免疫室，北京 100020; 2. 中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室，北京 102206

收稿日期: 2019-09-02

网络出版日期: 2019-04-25

基金资助

重大传染病防治国家科技重大专项(2018ZX10102001、2018ZX10711001、 2018ZX10734404), 传染病预防控制国家重点实验室发展基金项目(2011SKLID104)

收起

The change of endogenous siRNAs in coxsackie virus type B3-infected HeLa cells

YAO Hailan1 ,
SONG Juan2 ,
WANG Xinling2 ,
WANG Ruifang2 ,
SONG Qinqin2 ,
SHI Bintian2 ,
HAN Jun2

Expand

1. Department of Biochemistry & Immunology, Capital Institute of Pediatric, Beijing 100020, China; 2. State Key Laboratory of Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China

Received date: 2019-09-02

Online published: 2019-04-25

Fold

摘要

探究柯萨奇病毒B3型(Coxsackie virus type B3, CVB3)感染的细胞是否诱导内源性小干扰RNA(small interfering RNA，siRNA)的产生。以CVB3接种HeLa细胞，在细胞培养箱(5% CO2、37 ℃)孵育1 h。随后加入含2%血清的细胞维持液，继续培养3 h和6 h后收集细胞。用高通量测序(二代测序) 试剂盒提取细胞RNA，并反转录合成cDNA，构建文库，上机测序。过滤数据，去除插入片段过长的序列、低质量序列、poly A序列和小片段序列，与已知的小RNA数据库比对鉴定siRNA。通过茎-环反转录-聚合酶链反应，证实内源性siRNA在感染CVB3后的表达。结果显示，感染CVB3 3 h和6 h后，HeLa细胞产生了多种内源性siRNA。其中内源性siRNA(novel_sir3502和novel_sir2806)在感染后3 h和6 h均可持续表达。经比对，novel_sir3502和 novel_sir2806均可以识别45S和28S核糖体前体RNA。结果提示，CVB3感染可能干扰核糖体成熟。

关键词： 柯萨奇病毒B3型; HeLa细胞; 内源性小干扰RNA

本文引用格式

姚海兰1 , 宋娟2 , 王欣玲2 , 王瑞芳2 , 宋芹芹2 , 史冰田2 , 韩俊2 . 柯萨奇病毒B3型感染引起HeLa细胞内源性小干扰RNA的变化[J]. 微生物与感染, 2020 , 15(2) : 98 -103 . DOI: 10.3969/j.issn.1673-6184.2020.02.005

Abstract

To explore whether coxsackie virus type B3 (CVB3) infection can induce the expression of endogenous siRNA, siRNAs were analyzed in CVB3-infected HeLa cells at 3h and 6h post-infection (pi) by the high-throughput sequencing. The results showed that CVB3 induced 14 endogenous siRNA and 5 endogenous siRNA in HeLa cells at 3 h and 6 h pi, respectively. Two sustainably expressed endogenous siRNAs were further confirmed at 3 h and 6 h pi using quantitative PCR. Results of sequence homology alignment showed that two siRNAs could recognize the precursor RNA of 45S ribosome and 28S ribosome. The results suggest that CVB3 infection may interfere ribosome maturation.

Key words： Coxsackie virus type B3; HeLa cell; Endogenous siRNA

Options

文章导航

模态框（Modal）标题

摘要

本文引用格式

Abstract