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寨卡病毒感染人神经母细胞瘤细胞的转录组分析
姚秋凤,张力健,高雅琪,任浩,秦照玲*,戚中田*
0
(海军军医大学(第二军医大学)海军医学系生物医学防护教研室, 上海市医学生物防护重点实验室, 上海 200433
*通信作者)
摘要:
目的 通过生物信息学技术分析寨卡病毒(ZIKV)感染人神经母细胞瘤细胞SH-SY5Y后的转录组数据,拟找出参与ZIKV致病机制的潜在基因。方法 用ZIKV感染SH-SY5Y细胞,提取细胞总RNA后用转录组测序技术分析筛选出差异表达基因(DEG)。对DEG进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析,预测DEG主要参与的生物学过程、分子功能和信号通路,最后通过qPCR进行验证。结果 共鉴定出259个DEG,包括172个上调基因和87个下调基因。GO功能富集分析显示,DEG主要与细胞外基质、刺激应答、抗微生物体液反应、发育过程相关;KEGG通路富集分析显示,DEG主要与炎症反应、免疫反应有关。DEG的qPCR验证结果与转录组测序结果基本一致。结论 ZIKV感染SH-SY5Y细胞后参与细胞外基质、刺激应答、调控炎症反应的基因表达显著改变,说明ZIKV可能通过重塑细胞外基质及调控炎症反应引起神经系统病变。
关键词:  寨卡病毒  神经病变  高通量测序  转录组分析
DOI:10.16781/j.CN31-2187/R.20240149
投稿时间:2024-03-05修订日期:2024-05-07
基金项目:
Transcriptome analysis of human neuroblastoma cells infected with Zika virus
YAO Qiufeng,ZHANG Lijian,GAO Yaqi,REN Hao,QIN Zhaoling*,QI Zhongtian*
(Department of Biomedical Defense, Faculty of Naval Medicine, Naval Medical University (Second Military Medical University); Shanghai Key Laboratory of Medical Bioprotection, Shanghai 200433, China
*Corresponding authors)
Abstract:
Objective To analyze the transcriptome data of Zika virus (ZIKV)-infected human neuroblastoma cells SH-SY5Y by bioinformatics method, and to identify the potential genes involved in the pathogenic mechanism of ZIKV. Methods SH-SY5Y cells were infected with ZIKV.The total RNA was extracted and the differentially expressed genes (DEGs) were screened by transcriptome sequencing.Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to predict the biological processes, molecular functions, and signaling pathways mainly involved in the DEGs, and the results were verified by quantitative polymerase chain reaction (qPCR). Results A total of 259 DEGs were identified, including 172 up-regulated genes and 87 down-regulated genes.GO functional enrichment analysis showed that the DEGs were mainly related to extracellular matrix, response to stimulus, antimicrobial humoral response, and developmental process.KEGG pathway enrichment analysis revealed that the DEGs were predominantly associated to inflammatory reaction and immune response.The qPCR validation results of DEGs were basically consistent with the transcriptome sequencing results. Conclusion The expression of genes involved in extracellular matrix, response to stimulus, and regulation of inflammatory reaction is significantly altered in SH-SY5Y cells after ZIKV infection, suggesting that ZIKV may cause neurological lesions by remodeling the extracellular matrix and regulating inflammatory reaction.
Key words:  Zika virus  neuropathy  high-throughput sequencing  transcriptome analysis