论文部分内容阅读
人星状病毒(human astrovirus,HAstV)是引起婴幼儿病毒性腹泻的重要病原之一,但其致病机制及与宿主相互作用的数据非常有限。本研究旨在了解HAstV感染细胞后对宿主基因在转录水平上的影响。首先,利用临床分离HAstV毒株感染CaCo-2细胞,用实时定量反转录-聚合酶链反应(reverse transcriptasepolymerase chain reaction,RT-PCR)检测细胞培养上清液中的病毒总RNA,结果显示接种后9~24h病毒总RNA拷贝数明显增加。然后,利用转录组测序全面比较感染与未感染HAstV的CaCo-2细胞转录本,筛选两者间的差异表达基因;并应用KEGG信号通路富集性方法分析差异表达基因相关的信号通路。结果显示,差异基因富集在两条信号通路上(P<0.05),分别为轴突导向通路和Ras信号通路。本研究首次利用深度测序技术分析了HAstV感染对宿主基因在转录水平上的影响,为进一步研究HAstV致病机制等提供了方向。
Human astrovirus (HAstV) is one of the most important causative agents of viral diarrhea in infants and young children. However, its pathogenic mechanism and its interaction with host are very limited. The aim of this study was to understand the effect of HAstV on the transcriptional level of the host gene after it was infected. First, CaCO-2 cells were infected with clinically isolated HAstV strain and total viral RNA in the cell culture supernatant was detected by reverse transcriptase polymerase chain reaction (RT-PCR). The results showed that inoculation After 9 ~ 24h viral total RNA copy number increased significantly. Then, transcriptome sequencing was used to comprehensively compare the transcripts of CaCo-2 cells infected with and without HAstV, and to screen the differentially expressed genes between them. The signal pathways related to differentially expressed genes were analyzed by KEGG signaling pathway enrichment assay. The results showed that the differential genes were enriched in two signal pathways (P <0.05), which were the axon guidance pathway and the Ras signaling pathway, respectively. This study, for the first time, uses depth sequencing to analyze the effect of HAstV infection on the transcriptional level of host genes, which provides a direction for further study on the pathogenesis of HAstV.