MDCK细胞和MDCK-G1细胞感染H1N1流感病毒后病毒滴度和细胞转录组表达差异分析  

Analyzing the differences in virus titer and cell transcriptome expression between MDCK and MDCK-G1 cells infected with H1N1 influenza virus

作　　者：李兴航 杨柏峰 刘京 刘琛 张家友 杨晓明 LI Xing-hang;YANG Bai-feng;LIU Jing;LIU Chen;ZHANG Jia-you;YANG Xiao-ming(The Second Research Department of Virual Vaccines,Wuhan Institute of Biological Products Co.,Ltd.,Wuhan 430207,Hubei Province,China;不详)

机构地区：[1]武汉生物制品研究所有限责任公司病毒性疫苗研究二室,湖北武汉430207 [2]国家联合疫苗工程技术研究中心,湖北武汉430207 [3]中国生物技术股份有限公司,北京100029

出　　处：《微生物学免疫学进展》2021年第1期8-16,共9页Progress In Microbiology and Immunology

摘　　要：目的通过分析细胞表达基因及其相关信号通路的差异等信息,探索MDCK和MDCK-G1细胞株在接种H1N1流感病毒后出现病毒滴度和细胞生长趋势差异的内在生物学特性等原因。方法通过Illumina测序平台对2株细胞进行转录组测序(RNA sequencing,RNA-seq)和测序结果生物信息学分析后,选择了17个差异基因进行实时定量聚合酶链反应(quantitative real-time polymerase chain reaction,qRT-PCR)验证。结果MDCK和MDCK-G1细胞差异基因(differentially expressed genes,DEGs)|log_(2)(FoldChange)|>0和padj≤0.05总数为2786个。相比于MDCK细胞,MDCK-G1细胞有967个基因的表达显著上升,1819个基因的表达显著下降。差异基因通过基因本体(Gene Ontology,GO)数据库富集功能注释和京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)进行通路分析发现,2株细胞在免疫调控和细胞生长周期调控上均存在差异,qRT-PCR验证的17个基因中有16个基因的表达趋势与转录组测序结果一致。结论2株细胞的转录谱存在显著差异。Objective By analyzing information such as differences in cell expression genes and their associated signaling pathways,searching the intrinsic biological characteristics of differences in viral titer and cell growth trends occurred between MDCK and MDCK-G1 cell lines after being inoculated with H1 N1 influenza virus.Methods Two cell lines were sequenced by RNA sequencing(RNA-seq)on the Illumina platform.The sequencing results were analyzed by bioinformatics and 17 differentially expressed genes were selected for quantitative real-time polymerase chain reaction validation.Results The total number of differential genes(|log_(2)(FoldChange)|>0 and padj≤0.05)in MDCK and MDCK-G1 cells was 2786,of which 967 genes were significantly increased and 1819 genes were significantly decreased in MDCK-G1 cell compared to MDCK cell.The results showed that there were differences in cell growth,cell cycle regulation and immune regulation of influenza virus by analysis of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes database.The expression trend of 16 of the 17 genes validated by qRT-PCR was consistent with that of RNA-seq.Conclusion There are significant differences in the gene transcription spectrum between the two cell lines.

关 键 词：MDCK细胞 转录组学 流感病毒 差异 

分 类 号：R373.13[医药卫生—病原生物学]