人呼吸道合胞病毒感染不同细胞基因的差异表达分析  

Analysis of differentially gene expression in different cells infected with human respiratory syncytial virus

作　　者：万常青 程宁宁 苏彦斌 李辰霄 付远辉[2] 何金生[2] 高博 杜琳 WAN Changqing;CHENG Ningning;SU Yanbin;LI Chenxiao;FU Yuanhui;HE Jinsheng;GAO Bo;DU Lin(Recombinant Vector Vaccine R&D Center,Beijing Zhifei Luzhu Biopharmaceutical Co.,Ltd.,Beijing 100176,China;不详)

机构地区：[1]北京智飞绿竹生物制药有限公司重组载体疫苗研发中心,北京100176 [2]北京交通大学生命科学与生物工程研究院,北京100091

出　　处：《微生物学免疫学进展》2023年第5期11-16,共6页Progress In Microbiology and Immunology

摘　　要：目的 采用生物信息学方法,筛选人呼吸道合胞病毒(human respiratory syncytial virus,RSV)感染人喉表皮样癌细胞(human larynx epidermoid carcinoma cells,HEp-2)与人肺(支气管)上皮细胞(human normal lung epithelial cell,BEAS-2B)后均具有表达差异的基因,为了解宿主与RSV之间的相互作用提供线索。方法 从基因表达数据库(gene expression omnibus,GEO)中下载RSV感染HEp-2与BEAS-2B细胞的转录组数据,运用R软件进行转录组数据差异表达分析,筛选差异表达基因。利用DAVID数据库对差异表达基因进行基因本体(gene ontology,GO)功能富集和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路分析,采用STRING数据库网站构建蛋白共作网络(Protein-Protein Interaction Network Analysis,PPI),并运用Cytoscape软件中的CytoHubba插件寻找差异表达基因中的关键基因。最后,采用实时定量PCR(real-time quantitative PCR,qRT-PCR)对RSV感染HEp-2与BEAS-2B细胞后均差异表达的基因进行验证。结果 筛选出135个RSV感染HEp-2与BEAS-2B细胞后均差异表达的基因,其中有132个上调基因,3个下调基因。这些基因主要富集在炎症反应和肿瘤坏死因子信号通路。CytoHubba评分确定2个关键基因CTGF和IL-11,qRT-PCR结果显示,2种细胞被RSV感染后,CTGF与IL-11基因表达水平均显著增强。结论 本研究通过对基因转录组数据的分析以及实验验证,发现CTGF和IL-11基因在RSV感染的HEp-2与BEAS-2B细胞中均表达显著增强,与转录组数据差异表达分析结果一致,提示2个基因参与了RSV感染过程,CTGF和IL-11是影响RSV复制的宿主基因,该发现有助于找到防治RSV的新靶点,为RSV防治策略提供新思路。Objective Bioinformatics methods were utilized to screen for differently expressed genes in HEp-2 and BEAS-2B cells infected with human respiratory syncytial virus(RSV),providing insights into the interaction between host cells and RSV.Methods Transcriptomic data pertaining to RSV-infected HEp-2 and BEAS-2B cells were downloaded from the Gene Expression Omnibus(GEO) database.Employing the R software,an analysis of differential gene expression was executed with the objective of identifying genes that exhibited discernible alterations in their expression levels.The differentially expressed genes underwent gene ontology(GO) functional enrichment analysis and Kyoto Encyclopedia of Gene and Genomes(KEGG) pathway analysis,both of which were performed utilizing the DAVID database.The protein-protein interaction(PPI) network was constructed employing the STRING database.The CytoHubba plugin within Cytoscape was employed to identify key genes among the differentially expressed gene set.Finally,the differential expression of these genes was verified through real-time quantitative PCR(qRT-PCR) in RSV-infected HEp-2 and BEAS-2B cells.Results A total of 135 differentially expressed genes were identified following the screening of RSV-infected HEp-2 and BEAS-2B cells,with 132 genes showing upregulation and 3 genes showing downregulation.These genes were primarily enriched in inflammatory response and tumor necrosis factor signaling pathways.Based on the CytoHubba score,two key genes,CTGF and IL-11,were identified.The qRT-PCR results demonstrated a significant enhancement in the expression levels of CTGF and IL-11 genes in both cell types upon RSV infection.Conclusion This study conducted transcriptomic analysis and experimental validation to identify significant upregulation of CTGF and IL-11 genes in RSV-infected HEp-2 and BEAS-2B cells.These findings align with the differential expression analysis results,suggesting the involvement of these genes in RSV infection.CTGF and IL-11 are host genes influencing RSV replication,offe

关 键 词：人呼吸道合胞病毒 HEP-2细胞 BEAS-2B细胞 差异表达基因 生物信息学 转录组学 实时定量PCR 

分 类 号：R378[医药卫生—病原生物学]