猪圆环病毒2型感染人源细胞的转录组分析

Transcriptome analysis of human cells infected with porcine circovirus type 2

目的圆环病毒2型(porcine circovirus type 2,PCV2)能否感染人类细胞一直都是一个争论的话题。本研究通过转录组测序(RNA sequencing,RNA-seq)技术分析PCV2感染人源细胞后的差异表达基因及其相关的信号通路、生物学功能,以期为人源细胞对PCV2及其他病毒产生抗病毒效应的研究提供参考。方法用PCV2感染He La细胞,然后进行RNA-seq技术分析,筛选与抗病毒反应相关的差异表达基因(differentially expressed genes,DEGs),并通过实时定量PCR进一步验证。结果在感染PCV2的He La细胞中共注释到15402个Uni Genes,其中有387个DEGs(包括267个上调基因和120个下调基因)。对生物过程类DEGs进行GO富集和KEGG通路分析,发现富集基因要参与应激反应(97/291 DEGs)和免疫系统过程(80/291 DEGs),其中最显著两条途径的基因是NOD样受体信号途径(21/170 DEGs)和单纯疱疹病毒感染途径(22/170 DEGs)相关差异表达基因。结论 PCV2感染He La细胞之后,改变了细胞的表达谱,获得的DEGs及其功能注释信息有助于理解其抗病毒机制,对于了解PCV2感染人源细胞所产生的免疫反应及抗病毒效力提供依据。

Objective Whether porcine circovirus type 2(PCV2)can infect human cells is controversial.To study the antiviral effects of human cells on PCV2 and other viruses,we analyzed differentially expressed genes(DEGs)and their related signaling pathways and biological functions in PCV2-infected human cells by transcriptome analysis(RNA sequencing,RNA-seq).Methods HeLa cells were infected with PCV2,and then analyzed by RNA-seq to screen DEGs related to antiviral reactions.The DEGs were further verified by Real-time quantitative PCR.Results A total of 15402 UniGenes and 387 DEGs,including 267 upregulated genes and 120 downregulated genes,were annotated in human HeLa cells infected with PCV2.The most abundant Gene Ontology(GO)terms were response to stress(97/291 DEGs)and immune system process(80/291 DEGs).The most significant pathways were the nucleotide-binding and oligomerization domain(NOD)-like receptor signaling pathway(21/170 DEGs)and herpes simplex infection(22/170 DEGs).Conclusions The gene expression profile of HeLa cells significantly changed after PCV2 infection.The result obtained in this study will help understand the antiviral mechanism of human cells and may provide a basis for understanding the immune response and antiviral efficacy of human cells infected with PCV2.