Transcriptome sequencing analysis of SH-SY5Y cells infected with EV71 reveals the potential neuropathic mechanisms
Transcriptome sequencing analysis of SH-SY5Y cells infected with EV71 reveals the potential neuropathic mechanisms
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感染EV71的SH-SY5Y细胞的转录组测序分析揭示了潜在的神经病理机制
DOI：
10.1016/j.virusres.2020.197945
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发表时间：
2020-06-01
期刊：
影响因子：
5
通讯作者：
Zhang, Yunhui
中科院分区：
文献类型：
--
作者：
Hu, Yajie;Xu, Yanyan;Zhang, Yunhui
肠道病毒A71型（EV 71）是手足口病（HFMD）最常见的病原体，其引起的神经系统并发症通常是手足口病患儿死亡的主要原因。然而，EV 71感染引起神经系统改变的机制仍不清楚。因此，在本研究中，将EV 71接种到人神经母细胞瘤细胞系SH-SY 5 Y中，并随后使用转录组测序来检查感染的SH-SY 5 Y细胞中转录组的改变。它有望确定神经系统疾病响应EV 71感染的潜在机制。结果，在对照、EV 71 -12 h和EV 71 -24 h组中分别发现总共82，406，974、112，410，808和87，780，371个干净读段。此外，与对照组相比，在EV 71 -12 h和EV 71 -24 h组中分别鉴定出160和745个差异表达基因。接下来，为了进一步探讨EV 71感染引发的致病机制，我们主要集中在EV 71感染不同时间点的共同差异表达基因。发现了95个共同的差异表达基因，并将其用于GO和途径分析。GO富集分析划分了相关的生物学过程，分子功能和细胞组分，KEGG途径分析能够注释代谢途径并揭示显著富集的途径之间的相互作用。结果表明，丰富的GO术语“神经系统发育”和丰富的途径“CCKR信号通路图”可能是EV 71诱导的神经病理机制的重要贡献者。此外，我们还筛选了10个上调和下调的非蛋白质编码基因，这些基因在我们的转录组谱中表达显著不同，这表明这些异常调节的非蛋白质编码基因可能在EV 71感染的发病机制中也起重要作用。最终采用RT-qPCR技术对转录组测序数据进行验证，实验证明RT-qPCR和转录组测序结果基本一致。综上所述，这是首次对SH-SY 5 Y细胞对EV 71感染应答的转录组分析，为进一步探索EV 71感染引起的神经系统改变的机制提供了有价值的线索。
Enterovirus A71 (EV71) remains the most common causative agent of hand, foot, and mouth disease (HFMD), and the neurological complications induced by EV71 are usually the leading cause of death in children with HFMD. However, the mechanism of nervous system changes caused by EV71 infection is still unclear. Therefore, in the current study, EV71 was inoculated into the human neuroblastoma cell line SH-SY5Y and subsequent transcriptome sequencing was used to examine the alterations of the transcriptome in infected SH-SY5Y cells. It is expected to determine the underlying mechanism of neurological diseases in response to EV71 infection. As a result, a total of 82,406,974, 112,410,808 and 87,780,371 clean reads were found in the control, EV71-12 h and EV71-24 h groups, respectively. Moreover, 160 and 745 differentially expressed genes were identified in the EV71-12 h and EV71-24 h groups, respectively, as compared to the control group. Next, to further explore the pathogenic mechanism triggered by EV71 infection, we mainly focused on the common differentially expressed genes at different time points of EV71 infection. And it was discovered that there were 95 common differentially expressed genes, which were used to conduct GO and pathway analysis. GO enrichment analysis demarcated related biological processes, molecular functions and cellular components, and KEGG pathway analysis enabled annotations of metabolic pathways and revealed interactions among the significantly enriched pathways. The results showed that the enriched GO term "Nervous system development" and enriched pathway "CCKR signaling map" might be important contributors to EV71-induced neuropathological mechanisms. In addition, we also screened 10 up- and down-regulated non-protein coding genes with significantly different expression in our transcriptome profiling, which suggested that these abnormally regulated non-protein-encoding genes might also play important roles in the pathogenesis of EV71 infection. Eventually, RT-qPCR technology was adopted to validate the transcriptome sequencing data and the experiment demonstrated that the RT-qPCR and transcriptome sequencing results were basically consistent. In summary, this is the first transcriptome analysis of SH-SY5Y cells in response to EV71 infection and provides valuable cues for further exploring the mechanism of nervous system changes caused by EV71 infection.