ARRDC3在D68型肠道病毒感染A549细胞中的功能研究

李华怡; 傅慧超; 司俊卓; 杨春

doi:10.16016/j.2097-0927.202211078

您当前的位置：

首页 >

文章列表页 >

ARRDC3在D68型肠道病毒感染A549细胞中的功能研究

基础医学 | 更新时间：2026-01-09

- ARRDC3在D68型肠道病毒感染A549细胞中的功能研究
- Role of ARRDC3 in enterovirus D68-infected A549 cells
- 陆军军医大学学报 2023年45卷第14期页码：1539-1546
- 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：
  
  [ 杨春，E-mail: 100494@cqmu.edu.cn YANG Chun, E-mail: 100494@cqmu.edu.cn
- 基金信息：
  
  重庆市自然科学基金(CSTC2016jcyjA0212)
- DOI：10.16016/j.2097-0927.202211078
  中图分类号： R373.2;R394.3;R734.2
- 收稿：2022-11-11，
  
  修回：2023-04-23，
  
  纸质出版：2023-07-30
- 稿件说明：
移动端阅览
李华怡, 傅慧超, 司俊卓, 等. ARRDC3在D68型肠道病毒感染A549细胞中的功能研究[J]. 陆军军医大学学报, 2023,45(14):1539-1546.

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.
李华怡, 傅慧超, 司俊卓, 等. ARRDC3在D68型肠道病毒感染A549细胞中的功能研究[J]. 陆军军医大学学报, 2023,45(14):1539-1546. DOI： 10.16016/j.2097-0927.202211078.

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

SUN J, HU X Y, YU X F. Current understanding of human enterovirus D68[J]. Viruses, 2019, 11(6): 490. DOI: 10.3390/v11060490.

唐弘, 杨春, 康月茜, 等. 人肠道病毒D68型5'UTR间隔区域对下游基因表达的影响[J]. 微生物学通报, 2020, 47(12): 4196-4204. DOI: 10.13344/j.microbiol.china.200207.

TANG H, YANG C, KANG Y X, et al . Effect of EV-D68 virus 5'UTR spacer region on downstream gene expression[J ] . Microbiol China, 2020, 47(12): 4196-4204. DOI: 10.13344/j.microbiol.china.200207.

WANG X L, ZHANG P F, LI J, et al . Enterovirus D68 in a 6-year-old acute flaccid myelitis case in China, 2018: a case report[J ] . BMC Infect Dis, 2020, 20(1): 125. DOI: 10.1186/s12879-020-4829-y.

HARDY D, HOPKINS S. Update on acute flaccid myelitis: recognition, reporting, aetiology and outcomes[J]. Arch Dis Child, 2020, 105(9): 842-847. DOI: 10.1136/archdischild-2019-316817.

AYERS T, LOPEZ A, LEE A, et al . Acute flaccid myelitis in the United States: 2015-2017[J ] . Pediatrics, 2019, 144(5): e20191619. DOI: 10.1 542/peds.2019-1619.

SADEUH-MBA S A, JOFFRET M L, MAZITCHI A, et al . Genetic and phenotypic characterization of recently discovered enterovirus D type 111[J ] . PLoS Negl Trop Dis, 2019, 13(10): e0007797. DOI: 10.1371/journal.pntd.0007797.

李飞, 沈军, 谭文杰. 肠道病毒D组68型流行情况研究进展[J]. 中国病毒病杂志, 2020, 10(6): 471-475.

LI F, SHEN J, TAN W J. Recent progress in epidemiology of enterovirus D68[J]. Chin J Viral Dis, 2020, 10(6): 471-475.

SMITH J S, PACK T F, INOUE A, et al . Noncanonical scaffolding of G αi and β-arrestin by G protein-coupled receptors[J ] . Science, 2021, 371(6534): eaay1833. DOI: 10.1126/science.aay1833..

ZHANG Y H, LI M M, LI L Y, et al . β-arrestin 2 as an activator of cGAS-STING signaling and target of viral immune evasion[J ] . Nat Commun, 2020, 11(1): 6000. DOI: 10.1038/s41467-020-19849-9.

SMITH J S, PACK T F. Noncanonical interactions of G proteins and β-arrestins: from competitors to companions[J]. FEBS J, 2021, 288(8): 2550-2561. DOI: 10.1111/febs.15749.

WYLER E, MÖSBAUER K, FRANKE V, et al . Transcriptomic profiling of SARS-CoV-2 infected human cell lines identifies HSP90 as target for COVID-19 therapy[J ] . iScience, 2021, 24(3): 102151. DOI: 10.1016/j.isci.2021.102151.

TAKEUCHI F, KUKIMOTO I, LI Z Q, et al . Genome-wide association study of cervical cancer suggests a role for ARRDC3 gene in human papillomavirus infection[J ] . Hum Mol Genet, 2019, 28(2): 341-348. DOI: 10.1093/hmg/ddy390..

LEI D, DENG N, WANG S Q, et al . Upregulated ARRDC3 limits trophoblast cell invasion and tube formation and is associated with preeclampsia[J ] . Placenta, 2020, 89: 10-19. DOI: 10.1016/j.placenta.2019.10.009.

WEDEGAERTNER H, PAN W A, GONZALEZ C C, et al . The α-arrestin ARRDC3 is an emerging multifunctional adaptor protein in cancer[J ] . Antioxid Redox Signal, 2022, 36(13/14/15): 1066-1079. DOI: 10.1089/ars.2021.0193.

SHEN X, SUN X H, SUN B, et al . ARRDC3 suppresses colorectal cancer progression through destabilizing the oncoprotein YAP[J ] . FEBS Lett, 2018, 592(4): 599-609. DOI: 10.1002/1873-3468.12986.

ARAKAKI A K S, PAN W A, WEDEGAERTNER H, et al . α-Arrestin ARRDC3 tumor suppressor function is linked to GPCR-induced TAZ activation and breast cancer metastasis[J ] . J Cell Sci, 2021, 134(8): jcs254888. DOI: 10.1242/jcs.254888.

洪善超, 俞蕾, 黄嘉琪, 等. ARRDC3在前列腺癌中的表达及对前列腺癌细胞增殖、凋亡的影响[J]. 实用医学杂志, 2016, 32(21): 3582-3585. DOI: 10.3969/j.issn.1006-5725.2016.21.034.

HONG S C, YU L, HUANG J Q, et al . Expression of ARRDC3 in prostate cancer and its effect on proliferation and apoptosis of prostate cancer cells[J ] . J Pract Med, 2016, 32(21): 3582-3585. DOI: 10.3969/j.issn.1006-5725.2016.21.034.

SI J Z, TANG X, XU L, et al . High throughput sequencing of whole transcriptome and construct of ceRNA regulatory network in RD cells infected with enterovirus D68[J ] . Virol J, 2021, 18(1): 216. DOI: 10.1186/s12985-021-01686-x.

JHENG J R, CHEN Y S, HORNG J T. Regulation of the proteostasis network during enterovirus infection: a feedforward mechanism for EV-A71 and EV-D68[J]. Antivir Res, 2021, 188: 105019. DOI: 10.1016/j.antiviral.2021.105019.

ELRICK M J, PEKOSZ A, DUGGAL P. Enterovirus D68 molecular and cellular biology and pathogenesis[J]. J Biol Chem, 2021, 296: 100317. DOI: 10.1016/j.jbc.2021.100317.

陈暮霖, 穆星宇, 李维国, 等. 敲除含α-Arrestin结构域蛋白3对肾癌细胞PI3K/AKT信号通路及增殖能力的影响[J]. 现代生物医学进展, 2020, 20(13): 2401-2405. DOI: 10.13241/j.cnki.pmb.2020.13.001.

CHEN M L, MU X Y, LI W G, et al . Effects of α-arrestin-domain-containing-3 deficiency on PI3K/AKT signaling pathway and proliferation of renal cell carcinoma cells[J ] . Prog Mod Biomed, 2020, 20(13): 2401-2405. DOI: 10.13241/j.cnki.pmb.2020.13.001.

DORES M R, TREJO J. Endo-lysosomal sorting of G-protein-coupled receptors by ubiquitin: diverse pathways for G-protein-coupled receptor destruction and beyond[J]. Traffic, 2019, 20(2): 101-109. DOI: 10.1111/tra.12619.

SOUNG Y H, CHUNG H, YAN C, et al . Arrestin domain containing 3 reverses epithelial to mesenchymal transition and chemo-resistance of TNBC cells by up-regulating expression of miR-200b[J ] . Cells, 2019, 8(7): 692. DOI: 10.3390/cells8070692.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

鳖甲煎改良方下调CTGF/Itgα5对肝星状细胞T6周期变化、细胞凋亡及Col1a1表达的影响

Sox30基因敲除致小鼠睾丸间质细胞增殖异常的研究

HBx介导SAMHD1降解并通过Akt/p27通路促进肝癌细胞增殖的机制研究

基于基因芯片分析急性镉暴露致大鼠肝损伤及其可能机制

HBx基因通过ERK1/2信号通路促肝细胞增殖及炎性因子表达