埃可病毒11型感染RD细胞的转录组学分析及PLK1对其感染影响的初步探究  

Transcriptomics analysis of echovirus type 11 infected RD cells

作　　者：张国艳 李冀琛 张珂艺 孙强[1] 张勇[1] 武桂珍[1] Zhang Guoyan;Li Jichen;Zhang Keyi;Sun Qiang;Zhang Yong;Wu Guizhen(NHC Key Laboratory of Biosafety,NHC Key Laboratory of Medical Virology,World Health Organization Western Pacific Region Polio Reference Laboratory,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Weifang Medical University,Weifang 261053,China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,国家卫生健康委员会生物安全重点实验室,国家卫生健康委员会医学病毒与病毒病重点实验室,世界卫生组织西太平洋区脊髓灰质炎参比实验室,北京102206 [2]潍坊医学院,261053

出　　处：《中华实验和临床病毒学杂志》2021年第6期605-611,共7页Chinese Journal of Experimental and Clinical Virology

基　　金：北京市自然科学基金(L192014);国家自然科学基金(31900140);国家重点研发计划(2021YFC0863000);国家科技重大专项(2018ZX10734-401)。

摘　　要：目的探究埃可病毒11型(echovirus 11,E11)感染人横纹肌肉瘤细胞(human rhabdomyosarcoma cells,RD)后细胞内宿主因子在转录水平上的变化,初步了解Polo样激酶1(polo-like kinase 1,PLK1)对E11复制的影响及可能的分子机制。方法利用转录组测序(RNA-Seq技术)对E11感染RD细胞组和RD细胞对照组进行差异表达基因分析。使用基因本体论(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析显著差异表达基因,利用实时荧光定量聚合酶链式反应(quantitative real-time PCR,qPCR)对代表性差异基因的表达水平进行验证,并初步探究抑制PLK1对E11复制的影响。结果E11感染RD细胞后,共鉴定出3726个差异表达基因,感染组(感染后12 h、24 h和48 h)共同显著差异表达基因484个,这些显著差异表达基因主要富集在免疫反应、细胞内信号转导、转录因子活性、序列特异性DNA结合、蛋白激酶活性、核、膜的组成成分、PI3K-Akt信号通路等。使用qPCR对6个靶基因(CXCL14、IFITM1、OAS1、SAMD9、MX1和PLK1)的表达情况进行验证,均与RNA-Seq结果一致,其中CXCL14、IFITM1、OAS1、SAMD9、MX1表达上调倍数在1.2~104.97倍之间,PLK1表达下调倍数在0.99~5.79倍之间。抑制PLK1后,E11复制能力增加。结论炎症反应、PI3K-Akt、MAPK等通路可能在E11感染抗病毒免疫过程中发挥重要作用,此外,宿主因子PLK1可能通过调节细胞周期在E11感染中起到关键作用。Objective To investigate the transcriptional changes of host factors in human rhabdomyosarcoma cells(RD)infected with echovirus 11(E11),and to preliminarily understand the effect of polo like kinase 1(PLK1)on E11 replication and possible molecular mechanism.Methods Transcriptome sequencing(RNA SEQ)was used to analyze the differentially expressed genes between E11 infected RD cells group and RD cell control group.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)was used to enrich and analyze significantly differentially expressed genes.The expression situation of the representative differentially expressed genes were validated by quantitative real-time polymerase chain reaction(qRT-PCR),and the effect of PLK1 inhibition on E11 replication was preliminarily explored.Results A total of 3726 differentially expressed genes were identified in RD cells infected with E11,and 484 differentially expressed genes were found in the infected group(0 h,12 h,24 h and 48 h after infection).These differentially expressed genes were mainly enriched in the immune response,intracellular signal transduction,transcription factor activity,sequence-specific DNA binding,protein kinase activity,nuclear and membrane components,and PI3K-Akt signaling pathway.qPCR was used to verify the expression of six target genes(CXCL14,IFITM1,OAS1,SAMD9,MX1 and PLK1),which were consistent with result of RNA-Seq.The expression of CXCL14,IFITM1,OAS1,SAMD9,and MX1 were up-regulated by 1.2-104.97 times,and the expression of PLK1 was down regulated by 1-5.79 times.Inhibition of PLK1 increased E11 replication.Conclusions Inflammatory response,PI3K-Akt,MAPK and other pathways may play an important role in the antiviral immune process of E11 infection.In addition,host factor PLK1 may play a key role in E11 infection by regulating the cell cycle.

关 键 词：埃可病毒11型 转录组分析 POLO样激酶1 差异表达基因 宿主因子 

分 类 号：R373[医药卫生—病原生物学]