人呼吸道合胞病毒北京地区A亚型ON1基因型地方株感染A549细胞转录组学分析  

Transcriptomic analysis of A549 cells infected with ON1 genotype of human respiratory syncytial virus subtype A isolated in Beijing

作　　者：王凤杰 江明礼 郭琪 李晓芸 孙宇[1] 朱汝南[1] 赵林清[1] Wang Fengjie;Jiang Mingli;Guo Qi;Li Xiaoyun;Sun Yu;Zhu Runan;Zhao Linqing(Laboratory of Virology,Beijing Key Laboratory of Etiology of Viral Diseases in Children,Capital Institute of Pediatrics,Beijing 100020,China)

机构地区：[1]首都儿科研究所病毒学研究室,儿童病毒病病原学北京市重点实验室,北京100020

出　　处：《中华微生物学和免疫学杂志》2024年第2期128-133,共6页Chinese Journal of Microbiology and Immunology

基　　金：首都卫生发展科研专项(首发2022-1G-1131);北京市卫生健康委员会高层次技术人才建设(学科带头人-02-20)。

摘　　要：目的应用转录组测序方法,分析北京地区人呼吸道合胞病毒(respiratory syncytial virus,RSV)A亚型优势流行基因型ON1地方株感染A549细胞后差异表达基因,为RSV防治提供潜在靶点。方法选用已经过全基因组测序确定为RSV A亚型ON1基因型的地方株(61397-ON1)感染A549细胞,提取总mRNA,通过转录组测序筛选出与未感染的A549细胞为对照的差异表达基因,对其进行GO分析、KEGG通路分析,同时随机选择6个差异表达倍数大于2倍的基因进行qRT-PCR验证。结果以未感染的A549细胞为对照,筛选出1632个差异表达基因,其中807个基因表达上调,825个基因表达下调。差异基因主要参与细胞因子反应以及MAPK级联反应正向调控等免疫应答相关生物过程,并在MAPK信号通路、NOD样受体信号通路、p53信号通路、TNF信号通路、IL-17信号通路及NF-κB信号通路发生了富集。选择的6个差异表达基因qRT-PCR验证结果与转录组数据趋势一致。结论RSV A亚型ON1基因型毒株感染A549细胞后的差异表达基因主要参与细胞因子应答及免疫相关信号通路,为RSV致病的分子机制及防治策略研究提供基础资料。Objective To analyze the differentially expressed genes of human respiratory syncytial virus(RSV)subtype A genotype ON1,a predominant genotype in Beijing,after infecting A549 cells using transcriptomic sequencing,and provide potential targets for RSV prevention and treatment.Methods A local strain(61397-ON1)identified by whole-genome sequencing as ON1 genotype of RSV subtype A was selected to infect A549 cells.Total mRNA was extracted,and the differentially expressed genes in infected and uninfected A549 cells were screened by transcriptomic sequencing.GO analysis and KEGG pathway analysis were performed.Besides,six genes with differential expression ratio greater than two times were randomly selected for qRT-PCR verification.Results There were 1632 differentially expressed genes between infected and uninfected A549 cells,of which 807 genes were up-regulated and 825 genes were down-regulated.The differentially expressed genes were mainly involved in immune response-related biological processes such as cytokine response and positive regulation of MAPK cascades,and were enriched in MAPK signaling pathway,NOD-like receptor signaling pathway,p53 signaling pathway,TNF signaling pathway,IL-17 signaling pathway and NF-κB signaling pathway.The results of qRT-PCR for six differentially expressed genes were consistent with the trend of transcriptome data.Conclusions The differentially expressed genes of RSV A subtype ON1 genotype after infecting A549 cells are mainly involved in cytokine response and immune-related signaling pathways.This study provides basic data for further study of the molecular mechanism of RSV infection and the development of prevention and treatment strategies.

关 键 词：呼吸道合胞病毒 地方株 ON1基因型 转录组学分析 差异表达基因 

分 类 号：R56[医药卫生—呼吸系统]