基于RNA-seq分析戊型肝炎病毒感染HepG2细胞后的差异表达基因

Analysis of differently expressed genes of hepatitis E virus infected HepG2 cells using RNA-Seq technique

目的研究戊型肝炎病毒(Hepatitis E virus,HEV)感染人肝癌细胞HepG 2前后,HepG 2细胞的相关基因表达变化,为初步探索HEV在宿主细胞内感染机制及致病机理奠定基础。方法对HEV感染组HepG 2细胞和对照组HepG 2细胞的RNA进行高通量测序(RNA-seq技术),利用生物信息学方法对测序数据进行分析,对感染组和对照组的差异表达基因进行筛选、功能注释和通路富集分析。结果以基因表达差异倍数在2倍及以上为标准,从感染组与对照组共筛选出差异表达基因132个,其中,感染组相比于对照组,上调表达基因127个,下调表达基因5个。Gene Ontology(GO)功能富集分析注释结果显示,差异表达基因主要富集在病毒防御反应、固有免疫应答、病毒基因组复制的负调控及干扰素应答等过程;主要行使RNA结合、蛋白结合、调节解旋酶活性、NAD+ADP-核糖基转移酶的活性等分子功能。利用KEGG数据库作为参考,这些基因主要参与甲型流感、单纯疱疹感染、麻疹、C型肝炎、RIG-I样受体信号通路、病毒致癌、乙型肝炎、Toll样受体信号通路、胞质DNA传感通路、趋化因子信号转导通路和细胞凋亡等通路。结论通过功能及通路筛选,发现DDX58、STAT1、CXCL8、STAT2、TLR3、CXCL10、EIF2AK2、IRF9和IFIH1等基因可能在HEV感染抗病毒免疫过程中发挥重要作用。

To detect the changes in the expression of hepatitis E related genes in human HepG 2 cells infected by HEV,and to lay the foundation for the preliminary exploration of the infection mechanism and pathogenesis of HEV in the host cells.The HepG 2 cell RNA from the HEV infected group and control group were sequenced by high throughput sequencing(RNA-seq technology),and the analyzed by bioinformatics methods including screening,gene ontology functional enrichment analysis and pathway enrichment analysis of the differentially expressed genes in infected group and the control group.A total of 132 genes with at least 2-fold changes were identified from the infected group and the control group,of which 127 were up-regulated and 5 were down-regulated.The GO functional annotation shows that,the differentially expressed genes are mainly enriched in the process of defense response to virus,innate immune response,negative regulation of viral genome replication and response to interferon,and mainly play function of RNA binding,protein binding,regulation of the activity of helicase and the NAD+ADP-ribosyltransferase.Using the KEGG database as a reference,these genes are mainly involved in the pathway of influenza A,herpes simplex infection,measles,hepatitis C,RIG-I like receptor signaling pathway,virus carcinogenesis,hepatitis B,toll like receptor signaling pathway,cytoplasmic DNA-sensing pathway,chemokine signal transduction pathway and cell apoptosis.In conclusion through enrichment of functions and signaling pathways,it is found that DDX58,STAT1,CXCL8,STAT2,TLR3,CXCL10,EIF2AK2,IRF9 and IFIH1 may play an important role in the process of HEV infection.