基础医学与临床 ›› 2022, Vol. 42 ›› Issue (1): 68-74.

• 研究论文 • 上一篇    下一篇

柯萨奇病毒B组5型感染的人恶性胚胎横纹肌肉瘤细胞系lncRNA表达谱分析

滕培英,李静,师玉露,杨帆,欧霞,陈伟   

  1. 昆明理工大学医学院
  • 收稿日期:2021-05-18 修回日期:2021-11-03 出版日期:2022-01-05 发布日期:2022-01-05
  • 通讯作者: 陈伟 E-mail:55685836@qq.com
  • 基金资助:
    柯萨奇病毒B5调控宿主细胞天然免疫应答NF-κB信号通路 分子机制的研究

LncRNA expression profile in human malignant embryonic rhabdomyosarcoma cell line infected with Coxsackie virus group B type 5

  • Received:2021-05-18 Revised:2021-11-03 Online:2022-01-05 Published:2022-01-05
  • Contact: 0 00 E-mail:55685836@qq.com

摘要: 目的 探索柯萨奇病毒B组5型(CVB5)感染人恶性胚胎横纹肌肉瘤细胞系(RD)中差异长链非编码RNA(lncRNA)表达谱,为研究CVB5与宿主之间相互作用分子机制提供参考。方法 CVB5按感染复数为1接种RD细胞24 h后提取总RNA,采用转录组测序技术获取细胞中lncRNA差异表达谱,通过聚类分析、GO分析和KEGG通路富集对差异表达转录本进行了生物信息学分析。同时,利用RNAfold软件对lncRNA进行了二级结构预测。结果 与对照组相比,CVB5感染RD细胞后,共有1 754个mRNAs和508个lncRNA呈上调表达,3 106个mRNAs和760个lncRNA呈下调表达。差异表达lncRNA的共表达基因主要富集在分子结构活性、蛋白质分子结合和体液免疫反应等生物过程;lncRNA靶基因主要参与嗅觉转导途径、细胞因子受体相互作用和神经活性配体受体相互作用等通路。此外,实时荧光定量PCR验证的7个差异表达lncRNA与测序结果一致。结论 CVB5感染RD细胞后,差异显著性lncRNA主要参与了免疫相关过程,为充分理解lncRNA在CVB5感染中的调控作用奠定了基础。

关键词: RNA-seq, 柯萨奇病毒B组5型, 长链非编码RNA, RD细胞, 表达谱

Abstract: Objective: To explore the possible molecular mechanism of the interaction between Coxsackie virus Coxsackie virus group B type 5 (CVB5) and the host, the long non-coding RNA expression profile in human prototypic embryonal rhabdomyosarcoma cell line (RD) infected by CVB5 was investigated. Methods After 24 hours of RD cells infected with 1MOI CVB5 and extracted for RNA samples, and RNA-seq technology was used to obtain the differential expression profile of lncRNA in the cells, differentially expressed transcripts were analyzed by cluster analysis, GO analysis and KEGG pathway enrichment. At the same time, RNAfold was used to predict the secondary structure of lncRNA. Rusults Compared to the uninfected group, 1 754 mRNAs and 508 lncRNAs were up-regulated, and 3 106 mRNAs and 760 lncRNAs were down-regulated after CVB5 infection of RD cells. Co-expressed genes that differentially express lncRNA are mainly enriched in biological processes such as molecular structure activity, protein molecular binding, and humoral immune response; lncRNA targets are mainly involved in olfactory transduction pathway, cytokine-cytokine receptors interaction, neuroactive ligand-receptor interaction and other pathways. In addition, the seven differentially expressed lncRNAs verified by quantitative real-time PCR (RT-qPCR) are consistent with the sequencing results. Conclusions LncRNA is mainly involved in the regulation of the immune processes, which lays the foundation for a full understanding of the regulatory role of lncRNA in CVB5 infection .

Key words: RNA-seq, Coxsackie virus B5, long non-coding RNA, RD cells, expression profile