Role of ARRDC3 in enterovirus D68-infected A549 cells

Huayi LI; Huichao FU; Junzhuo SI; Chun YANG

doi:10.16016/j.2097-0927.202211078

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Role of ARRDC3 in enterovirus D68-infected A549 cells

Basic Medicine | 更新时间：2026-01-09

- Role of ARRDC3 in enterovirus D68-infected A549 cells
- Journal of Army Medical University Vol. 45, Issue 14, Pages: 1539-1546(2023)
- 作者机构：
  
  400016 重庆，重庆医科大学基础医学院病原生物学教研室
- 作者简介：
- 基金信息：
  
  the Natural Science Foundation of Chongqing(CSTC2016jcyjA0212)
- DOI：10.16016/j.2097-0927.202211078
  CLC： R373.2;R394.3;R734.2
- Received：11 November 2022，
  
  Revised：2023-04-23，
  
  Published：30 July 2023
- 稿件说明：
移动端阅览
Huayi LI, Huichao FU, Junzhuo SI, et al. Role of ARRDC3 in enterovirus D68-infected A549 cells[J]. Journal of Army Medical University, 2023, 45(14): 1539-1546.
DOI：

Huayi LI, Huichao FU, Junzhuo SI, et al. Role of ARRDC3 in enterovirus D68-infected A549 cells[J]. Journal of Army Medical University, 2023, 45(14): 1539-1546. DOI： 10.16016/j.2097-0927.202211078.

摘要

目的

探索含a-Arrestin结构域蛋白3(a-Arrestin-domain-containing-3，ARRDC3)在肠道病毒D组68型(enterovirus D species，EV-D68)感染A549中的表达以及对A549细胞株生物学行为的影响。

方法

对EV-D68感染宿主细胞前后的样本进行全转录组高通量测序分析，通过生物信息学分析筛选出差异表达基因ARRDC3；RT-qPCR和Western blot验证ARRDC3的表达水平；设计实验分组4组，对照组(未感染EV-D68的A549细胞)、感染组(感染EV-D68的A549细胞)、敲低ARRDC3感染组(感染EV-D68的A549细胞转染ARRDC3-SiRNA-干扰片段)、敲低对照感染组(感染EV-D68的A549细胞转染NC-SiRNA-干扰片段)，通过流式细胞术、CCK-8、Transwell实验检测ARRDC3在EV-D68感染模型中对A549细胞周期、增殖、迁移的影响。

结果

从测序结果中共筛选出差异表达2倍以上的差异表达基因(differentially expressed genes，DEGs)239个，其中上调基因135个，下调基因104个，ARRDC3是差异表达基因之一。EV-D68感染A549后，通过RT-qPCR和Western blot结果发现ARRDC3的表达水平显著增加(

0.05)。细胞周期明显缩短，抑制细胞迁移。进一步通过转染成功干扰ARRDC3表达载体，沉默ARRDC3可以增强EV-D68感染A549细胞，沉默ARRDC3细胞周期DNA合成期延长(

0.05)，虽然对宿主细胞增殖无显著影响，但沉默ARRDC3可引起细胞迁移能力增强(

0.05)。

结论

ARRDC3在EV-D68感染A549后表达明显上调，通过阻滞宿主细胞DNA合成期，抑制宿主细胞迁移等作用来发挥对宿主细胞的影响。

Abstract

Objective

To explore the expression of a-arrestin domain containing protein 3 (ARRDC3) in EV-D68 infected A549 cells and its effect on the biological behaviors of the cells.

Methods

The samples before and after EV-D68 infecting host cells (A549 cells) were analyzed by high-throughput whole transcriptome analysis with sequencing. After bioinformatics analysis was used to screen differentially expressed genes (DEGs)

RT-qPCR and Western blotting were used to verify the expression of ARRDC3 at mRNA and protein levels. Then the A549 cells were designed and divided into 4 groups: control group (A549 cells not infected with EV-D68)

infection group (A549 cells infected)

knockdown ARRDC3 infection group (ARRDC3 siRNA interference-mediated knockdown in A549 cells after infection)

knockdown control infection group (A549 cells infected with EV-D68 transfected with NC siRNA interference). Flow cytometry

CCK-8 assay

and Transwell assay were used to determine the effects of ARRDC3 on cell cycle

proliferation

and migration in above groups of cells.

Results

Gene sequencing showed there were 239 DEGs with more than 2-fold differential expression

with 135 up-regulated and 104 down-regulated genes

and ARRDC3 is one of DEGs. After A549 cells were infected with EV-D68

the expression level of ARRDC3 was significantly increased through RT-qPCR and Western blotting (

0.05)

cell cycle was arrested and cell migration was inhibited. Silencing of ARRDC3 through siRNA transfection improved EV-D68 infection to A549 cells and prolonged the cell cycle arrested at DNA synthesis phase (

0.05). Although the silencing showed no significant effect on the proliferation of host cells

cell migration ability was obviously enhanced (

0.05).

Conclusion

ARRDC3 is significantly up-regulated after EV-D68 infects A549 cells

and exerts its influence on host cells by blocking the DNA synthesis phase and inhibiting migration of host cells.

关键词

Keywords

references

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