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APOBEC3G对HBV复制的影响及转录组学分析

Effect of APOBEC3G on HBV replication and transcriptomic analysis
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摘要 目的:基于转录组测序和生物信息学分析对(APOBEC)亚家族(APOBEC3G A3G)参与抑制乙型肝炎病毒(HBV)复制的潜在分子机制及下游信号通路预测。方法:将A3G与HBV的表达质粒(A3G组)、GFP与HBV的表达质粒(HBV组)以及空载质粒(Vector组)分别转染于Huh7细胞中,3 d后收集细胞并提取RNA进行转录组测序,检测样本中mRNA的表达情况。采用DESeq软件对基因的表达水平进行差异分析筛选出差异表达基因,通过对差异基因进行GO富集分析和KEGG通路富集分析,确定差异基因行使的主要功能和参与的通路,同时依据STRING数据库进行蛋白质-蛋白质互相作用(PPI)分析,构建蛋白互作网络,以寻找目的基因之间的相互关系及关键节点蛋白。结果:A3G组和HBV组相比,筛选出21个显著差异基因;GO富集分析显示的项目包括脂蛋白B mRNA编辑酶复合物的形成、EH结构域的结合、中胚层细胞命运规范的BMP信号传导途径等;KEGG通路富集分析显示可能与内吞作用和坏死相关;PPI网络发现了NOXA1、LSM10、RNF103-CHMP3等5个关键蛋白及其互作蛋白。HBV组和Vector组相比,筛选出42个显著差异基因;GO富集分析显示的项目包括离子通道活性、多种酶活性和膜成分等相关;KEGG通路富集分析显示与多种物质的代谢反应和p53信号传导等相关;PPI网络发现了UBD、CCND3、RHOD等15个关键蛋白及其互作蛋白。结论:通过转录组测序和生物信息学分析,探究A3G抑制HBV复制的潜在下游分子,丰富了A3G抑制HBV复制的机制研究。 Objective:To explore and predict the underlying mechanisms and downstream signaling pathway prediction of APOBEC3G(A3G)inhibiting Hepatitis B virus(HBV)based on transcriptome sequencing and bioinformatics analysis.Methods:The expression plasmid of A3G and HBV(A3G group),expression plasmid of GFP and HBV(HBV group)and empty plasmid(Vector group)were transfected into the Huh7 cells,respectively.Three days later,the cells were collected,and RNA was extracted for transcriptome sequencing to detect mRNA expression in the samples.DESeq software was used to conduct differential analysis on gene expression levels,and screen out differentially expressed genes.Through GO enrichment analysis and KEGG pathway enrichment analysis of differential genes,the main functions and pathways involved in differential genes were determined.Meanwhile,protein-protein interaction(PPI)analysis was conducted based on STRING database.The protein interaction network was constructed to find the relationship between the target genes and key node proteins.Results:Compared with the HBV group,21 significantly different genes were screened in A3G group.The results of GO enrichment analysis showed the item included the formation of lipoprotein B mRNA editing enzyme complex,EH domain binding and BMP signaling pathway regulated by mesoderm cell fate.KEGG pathway enrichment analysis showed that it might be related to endocytosis and necrosis.The PPI network identified NOXA1,LSM10,RNF103 and their interacting proteins.Compared with the Vector group,42 significantly different genes were screened in the HBV group.GO enrichment analysis shows projects related to the ion channel activity,various enzymatic activities,and membrane components,etc.KEGG pathway enrichment analysis shows associations with metabolic reactions of various substances and the p53 signaling pathway,and so on.The PPI network identified 15 key proteins such as UBD,CCND3 and RHOD,and their interacted proteins.Conclusions:Through transcriptome sequencing and bioinformatics analysis,the potential downstream molecules of A3G inhibiting HBV replication were explored,enriching the research on the mechanism of A3G′s inhibition of HBV replication.
作者 周未平 刘光焱 ZHOU Weiping;LIU Guangyan(Department of Pathogenic Biology,Shenyang Medical College,Shenyang Liaoning 110034;Liaoning Province Key Lab of Environmental Pollution and Microecology,Shenyang Liaoning 110034,China)
出处 《蚌埠医学院学报》 CAS 2024年第11期1411-1416,共6页 Journal of Bengbu Medical College
基金 国家自然科学基金资助项目(81702856) 辽宁省沈阳市中青年科技创新人才支持计划项目(RC210215) 辽宁省教育厅科学研究经费项目(青年项目)(LJKQZ2021176) 辽宁省科技计划面上项目(2022-MS-409) 沈阳医学院硕士研究生科技创新基金项目(Y20220508)。
关键词 乙型肝炎病毒 载脂蛋白B mRNA编辑酶催化多肽样蛋白 转录组测序 生物信息学分析 hepatitis B virus apolipoprotein B mRNA editing enzyme catalytic polypeptide like transcriptome sequencing bioinformatics analysis
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