肠道病毒D68型感染前后转录组分析及差异表达基因的验证

唐霞; 司俊卓; 卢楠; 何永林; 徐蕾; 包嘉佳; 唐佳玲; 杨春

doi:10.16016/j.1000-5404.202009104

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肠道病毒D68型感染前后转录组分析及差异表达基因的验证

基础医学 | 更新时间：2026-01-09

- 肠道病毒D68型感染前后转录组分析及差异表达基因的验证
- Transcriptome analysis and verification of differentially expressed genes before and after enterovirus D68 infection
- 第三军医大学学报 2021年43卷第4期页码：275-282
- 作者机构：
  
  400016 重庆，重庆医科大学基础医学院病原微生物学教研室
- 作者简介：
  
  [ 杨春，E-mail：yangchunim@163.com YANG Chun, E-mail:yangchunim@163.com
- 基金信息：
  
  重庆市自然科学基金(CSTC2016jcyjA0212)
- DOI：10.16016/j.1000-5404.202009104
  中图分类号： R373.2;R394-33;R394.3
- 收稿：2020-09-13，
  
  修回：2020-11-25，
  
  纸质出版：2021-04-15
- 稿件说明：
移动端阅览
唐霞, 司俊卓, 卢楠, 等. 肠道病毒D68型感染前后转录组分析及差异表达基因的验证[J]. 第三军医大学学报, 2021,43(4):275-282.

Xia TANG, Junzhuo SI, Nan LU, et al. Transcriptome analysis and verification of differentially expressed genes before and after enterovirus D68 infection[J]. Journal of Third Military Medical University, 2021, 43(4): 275-282.
唐霞, 司俊卓, 卢楠, 等. 肠道病毒D68型感染前后转录组分析及差异表达基因的验证[J]. 第三军医大学学报, 2021,43(4):275-282. DOI： 10.16016/j.1000-5404.202009104.

Xia TANG, Junzhuo SI, Nan LU, et al. Transcriptome analysis and verification of differentially expressed genes before and after enterovirus D68 infection[J]. Journal of Third Military Medical University, 2021, 43(4): 275-282. DOI： 10.16016/j.1000-5404.202009104.

摘要

目的

探讨肠道病毒D68型（enterovirus D68

EV-D68)感染前后宿主细胞转录组的表达变化，寻找与EV-D68感染的相关基因及信号通路，为初步探索EV-D68的感染机制奠定基础。

方法

将病毒以感染复数（multiplicity of infection，MOI）0.01接种于人横纹肌瘤（rhabdomyosarcoma

RD）细胞，24 h后提取感染组和对照组细胞总RNA进行转录组测序，筛选差异表达基因(differentially expressed genes

DEGs)，对其进行GO和KEGG富集分析，并用RT-qPCR验证8个主要差异表达的基因（

EGR

1、

fos

、

ZNF

625、

ARRDC

3、

jun

、

ASNS

、

CHAC

1、

ADM

2）。

结果

从测序结果中共筛选出差异表达两倍以上的基因140个，其中上调基因71个，下调基因69个。GO功能富集分析结果主要涉及离子的调控与应答；KEGG富集分析结果主要与甲状旁腺激素的合成分泌和作用、GnRH信号通路、Toll样受体信号通路、IL-17信号通路和MAPK信号通路等有关。RT-qPCR验证结果表明选取的8个基因上调或下调的差异表达倍数趋势与测序结果基本一致，

fos

和

jun

的表达量随病毒MOI及感染时长的增加而增加。

结论

通过对差异表达基因的功能和信号通路富集分析及验证，EV-D68加重哮喘的机制可能与

fos

上调有关；而MAPK信号通路可能在EV-D68感染期间发挥促炎作用和抗病毒作用。

Abstract

Objective

To detect the alterations of the transcriptome in host cells infected by enterovirus D68 (EV-D68) and look for the genes involved in its infection so as to lay a foundation for the preliminary exploration of the infection mechanism.

Methods

The virus was inoculated into human rhabdomyosarcoma (RD) cells at a multiplicity of infection (MOI) of 0.01. After 24 h

RNA was extracted from the infected RD cells and control cells for transcriptome sequencing. Differentially expressed genes (DEGs) were screened and analyzed by using Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Eight DEGs

EGR

fos

ZNF

625

ARRDC

jun

ASNS

CHAC

1 and

ADM

2 were validated by RT-qPCR.

Results

A total of 140 genes with at least 2-fold changes at expression level were identified from the sequencing results

of which 71 were up-regulated and 69 were down-regulated. The results of GO functional enrichment analysis mainly involved the regulation and response to ions while the results of KEGG enrichment analysis mainly related to parathyroid hormone synthesis

secretion and action

IL-17 and GnRH signaling pathway

Toll-like receptor signaling pathway

and MAPK signaling pathway and so on. The RT-qPCR results showed that the trend of fold change of expression in the selected 8 genes was basically consistent with the sequencing results

and the expression levels of

fos

and

jun

were increased with the increase of virus MOI and infection duration.

Conclusion

By validating and using the functional and enrichment analysis

we infer that the mechanism of EV-D68 exacerbating asthma may be related to the up-regulation of

fos

and MAPK signaling pathway may play a pro-inflammatory and antiviral role during EV-D68 infection.

关键词

Keywords

references

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