DENV-2感染HUVECs的基因表达谱分析及ceRNA调控网络的构建  

Analysis of gene expression profile and construction of ceRNA regulatory network after DENV-2 infection with HUVECs

作　　者：姬进忠 陈铭勰 胡盼 程瑶 王远迎 孙见飞 吴宁[1] 左丽[2] JI Jinzhong;CHEN Mingxie;HU Pan;CHENG Yao;WANG Yuanying;SUN Jianfei;WU Ning;ZUO Li(Chemistry and Biochemistry Laboratory,Guizhou Medical University,Guiyang 550025;Department of Immunology,Guizhou Medical University,Guiyang 550000,China)

机构地区：[1]贵州医科大学化学与生物化学实验室,贵州贵阳550025 [2]贵州医科大学免疫学教研室,贵州贵阳550000

出　　处：《西安交通大学学报（医学版）》2023年第4期532-541,共10页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81860289)。

摘　　要：目的旨在建立共表达lncRNA-mRNA的ceRNA网络,探究lncRNA在登革热中的潜在分子机制。方法通过基因芯片技术对DENV-2感染的和正常的pHUVEC进行测序并筛选出差异表达的lncRNA和mRNA。差异表达的mRNA进行蛋白互作(PPI)分析,利用Pearson相关系数筛选出显著相关的共表达lncRNA-mRNA,通过数据库预测能与共表达lncRNA-mRNA结合的miRNA,采用Cytoscape软件构建共表达lncRNA-mRNA的ceRNA网络。将差异表达的mRNA和共表达lncRNA-mRNA进行GO和KEGG富集分析,并通过RT-qPCR验证共表达lncRNA-mRNA。结果DENV-2感染48、72 h后,获得差异表达的mRNAs分别为105、51个,lncRNAs分别为59、29个;两个时间段共有差异mRNA 10个,lncRNA 5个。差异mRNAs进行PPI分析,得出IL6、IFIT2、OAS2等度值较高;lncRNA-mRNA共表达分析结果中,48 h和72 h相关系数最高的配对分别为XLOC_001966-SMTNL1和XLOC_001966-ESR2;GO和KEGG富集分析显示,差异表达的mRNA和共表达lncRNA-mRNA的功能主要集中在病毒防御反应、免疫应答、信号转导等,并通过JAK-STAT信号通路、Ⅰ型干扰素、细胞因子受体相互作用等途径发挥作用。RT-qPCR结果显示,共表达lncRNA-mRNA对中lncRNA XLOC-I2-8991上调,其余lncRNA和mRNA均下调。结论本研究初步揭示了登革病毒感染过程中潜在lncRNA-mRNA共表达网络,发现了共表达lncRNA-mRNA主要富集在病毒感染过程中的免疫调控及信号转导等途径,这将有助于进一步探索DENV-2的感染机制。Objective To establish a co-expression lncRNA-mRNA ceRNA network and explore the potential molecular mechanism of lncRNA in dengue fever.Methods DENV-2-infected and normal pHUVEC were sequenced and screened for differentially expressed lncRNA and mRNA by gene microarray technology.Differentially expressed mRNA was analyzed by protein-protein interaction(PPI),and significantly related co-expressed lncRNA-mRNA was screened by Pearson’s correlation coefficient.The microRNA(miRNA)that bound to co-expressed lncRNA-mRNA was predicted by the database.The ceRNA network of co-expressed lncRNA-mRNA was constructed by Cytoscape software.Finally differentially expressed mRNAs and co-expressed lncRNA-mRNA were analyzed by GO and KEGG enrichment,and co-expressed lncRNA-mRNA was verified by RT-qPCR.Results At 48 h and 72 h after infection,105 and 51 differentially expressed mRNAs were obtained,respectively,while 59 and 29 differentially expressed lncRNAs were obtained,respectively.Furthermore,at the two time intervals,there were 10 differential mRNAs and 5 differential lncRNAs,respectively.PPI analysis of differential mRNAs showed that isocratic values of interleukin 6(IL6),interferon-induced protein with tetratricopeptide repeats 2(IFIT2),and 2-5-oligoadenylate synthetase 2(OAS2)were relatively high.The pairing results of lncRNA-mRNA co-expression analysis with the highest correlation coefficients at 48 h and 72 h after infection were XLOC_001966-SMTNL1 and XLOC_001966-ESR2,respectively.According to Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,the functions of differentially expressed mRNA and co-expressed lncRNA-mRNA were mainly involved in virus epidemic prevention response,immune response,and signal transduction,as well as the Janus kinase(JAK)-signal transducer and activator of transcription(STAT)signaling pathway,type I interferon,and cytokine receptor interaction.RT-qPCR revealed that lncRNA XLOC-I2-8991 was upregulated in the co-expressed lncRNA-mRNA,whereas all the other lncRN

关 键 词：DENV-2 登革热 lncRNA MRNA CeRNA 

分 类 号：R392.12[医药卫生—免疫学]