基于Label-free定量蛋白组学揭示肠道病毒A71型感染致人扁桃体上皮细胞蛋白质组的改变

Label-Free-Based Quantitative Proteomics Reveals the Proteomic Alterations of Human Tonsillar Epithelial Cells Infected with Enterovirus-A71

肠道病毒A组71型(Enterovirus-A71,EV-A71)感染引起的手足口病给婴幼儿造成严重的疾病负担,明确EV-A71的致病机制能为有效防控手足口病提供科学依据。为了研究EV-A71感染与人扁桃体上皮细胞的互作,本研究采用Label-free定量蛋白组学技术研究EV-A71感染人扁桃体上皮细胞UT-SCC-60B 6h和12h后蛋白质组的改变情况,并确定EV-A71感染动力学(EV-A71 6h∶12h)过程中蛋白质组的改变情况,并进行差异表达蛋白(Differentially expressed proteins,DEPs)的蛋白互作网络和基因本体(Gene ontology,GO)分析。EV-A71感染6h时共有27个DEPs(下调12个DEPs,上调15个DEPs)发生显著改变;EV-A71感染12h时共有62个DEPs(下调26个DEPs,上调36个DEPs)发生显著改变;EV-A71感染动力学(EV-A71 6h∶12h)过程中共有40个DEPs(下调21个DEPs,上调19个DEPs)发生显著改变。显著发生改变的COX6C、TIMM10、MAP1LC3B、HIST1H1D、DNAJC8、TMED4和ARPC5等DEPs为各组间重叠蛋白。蛋白互作网络和GO分析结果显示,DEPs主要涉及到线粒体及线粒体膜的通透性调节、细胞囊泡形成、细胞凋亡、线粒体介导的凋亡等生物学过程。因此,EV-A71感染使人扁桃体上皮细胞的蛋白表达谱发生改变。

Hand,foot,and mouth disease(HFMD)is caused by enterovirus A71(EV-A71). HFMD poses a heavy disease burden upon young children. Elucidating the pathogenesis of EV-A71 infection could provide scientific evidence for prevention and control of the EV-A71. We wished to study the interaction between EVA71 infection and human tonsillar epithelial cells. We used label-free quantification proteomics to determine the proteomic alterations in a human tonsillar epithelial line(UT-SCC-60B)6 hrs,12 hrs and a dynamic process(6 h∶12 h)after EV-A71 infection. Differentially expressed proteins(DEPs)were analyzed through interaction networks and analysis of the Gene Ontology(GO) database. Expression of 27 DEPs(12 DEPs were downregulated and 15 DEPs were upregulated) was altered significantly 6 hrs after EV-A71 infection.Expression of 62 DEPs(26 DEPs were downregulated and 36 DEPs were upregulated)or 40 DEPs(21 DEPs were downregulated and 19 DEPs were upregulated)was altered significantly at 12 hrs or in a dynamic process(6 h∶12 h)after EV-A71 infection. DEPs such as COX6 C,TIMM10,MAP1 LC3 B,HIST1 H1 D,DNAJC8,TMED4 and ARPC5 were co-detected DEPs among three groups. The analysis of DEPs mainly referred to mitochondria and regulation of mitochondrial permeability,formation of vesicle organization,apoptosis and mitochondria-mediated apoptosis through a protein-interaction network and analyses of the GO database. In conclusion,the proteome of UT-SCC-60 B was altered by EV-A71 infection.