微生物与感染
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微生物与感染  2016, Vol. 11 Issue (2): 79-86    
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人星状病毒临床分离株感染CaCo­-2细胞的转录组分析
吴立梦1,2,*,滕峥1,*,刘晶2,张曦1,崔晓娴2,林庆能1,吴寰宇1,袁政安1,谢幼华2
1. 上海市疾病预防控制中心,上海 200336;2. 复旦大学基础医学院教育部/卫生部医学分子病毒学重点实验室,上海 200032
Transcriptome analysis of clinically isolated human astrovirus in CaCo­-2 cells
WU Li-Meng1,2,*, TENG Zheng1,*, LIU Jing2, ZHANG Xi1, CUI Xiaoxian2, LIN Qingneng1, WU Huanyu1, YUAN Zheng‘an1, XIE Youhua2
1. Shanghai Municipal Center for Disease Control and Prevention, Shanghai 200336, China; 2. Key Laboratory of Medical Molecular Virology of Ministries of Education and Health, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China
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摘要: 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9~24 h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。

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吴立梦1
2
*
滕峥1
*
刘晶2
张曦1
崔晓娴2
林庆能1
吴寰宇1
袁政安1
谢幼华2
关键词 人星状病毒转录组测序转录组    
Abstract:Human astrovirus (HAstV) is one of the major causes of viral gastroenteritis in infants and young children. However, data on the host response to and pathogenesis of HAstV infection are limited. The primary goal of this study was to identify differentially expressed genes and enriched pathways associated with HAstV infection in human CaCo­-2 cells. The clinical isolates of HAstV were used to infect CaCo-­2 cells and total viral RNAs in the extracellular medium were quantitatively detected. Real­time reverse transcriptase­polymerase chain reaction (RT­-PCR) assay was applied to clearly demonstrate a significant increase in the amount of viral RNAs in the culture supernatant at 9-­24 h post­infection. Finally, RNA sequencing was performed to compare the transcriptomes of CaCo­-2 cells infected with and without a clinically isolated HAstV strain. Statistical analyses on differentially expressed genes revealed that axon guidance and Ras signaling pathways were altered in HAstV­5-­infected CaCo­-2 cells (P<0.05). This is the first report on the deep­-sequencing analysis of transcripts of CaCo-­2 cells infected with HAstV. The data from this study can serve as a basis for future studies of HAstV pathogenesis.

Key wordsHuman astrovirus    RNA sequencing    Transcriptome
基金资助:“十二五”国家科技重大专项(2012ZX10004-211-03),上海市第四轮公共卫生三年行动计划重点学科建设(15GWZK0101)

通讯作者: 谢幼华   
引用本文:   
吴立梦1,2,*,滕峥1,*,刘晶2,张曦1,崔晓娴2,林庆能1,吴寰宇1,袁政安1,谢幼华2. 人星状病毒临床分离株感染CaCo­-2细胞的转录组分析[J]. 微生物与感染, 2016, 11(2): 79-86.
WU Li-Meng1,2,*, TENG Zheng1,*, LIU Jing2, ZHANG Xi1, CUI Xiaoxian2, LIN Qingneng1, WU Huanyu1, YUAN Zheng‘an1, XIE Youhua2. Transcriptome analysis of clinically isolated human astrovirus in CaCo­-2 cells. JOURNAL OF MICROBES AND INFECTIONS, 2016, 11(2): 79-86.
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http://jmi.fudan.edu.cn/CN/     或     http://jmi.fudan.edu.cn/CN/Y2016/V11/I2/79

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