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人巨细胞病毒感染对THP-1细胞自噬的影响。

The influence of HCMV infection on autophagy in THP-1 cells.

作者信息

Liu Yujie, Pan Jianwei, Liu Lifang, Li Wei, Tao Ran, Chen Yinghu, Li Huamei, Shang Shiqiang

机构信息

Department of Central Laboratory, Children's Hospital of Zhejiang University School of Medicine, Hangzhou, China.

出版信息

Medicine (Baltimore). 2017 Nov;96(44):e8298. doi: 10.1097/MD.0000000000008298.

Abstract

BACKGROUND

Human cytomegalovirus (HCMV) infection is very common and latency can be reactivated in the future. And it can alter the intracellular environment, similar to other herpesviruses, for viral replication and survival. The aim of this study was to investigate the influence of HCMV infection on autophagy in human acute monocytic leukemia cell line (THP-1 cells).

METHODS

Reverse transcription polymerase chain reaction (RT-PCR), western blot, and transmission electron microscope (TEM) were used to examine autophagy level. The concentrations of autophagy-related proteins Beclin 1, Atg5, and the light chain three (LC3) were counted when compared with actin level.

RESULTS

The expression levels of Beclin1, Atg5, and LC3II mRNAs increased gradually between 1 and 5 days(d) postinfection (p.i.) and subsequently decreased little by little when compared with the control THP-1 cells. However, results of western blot analysis displayed that the level of LC3II increased gradually after 1 day p.i. and decreased at 7 days after infection. But the levels of Atg5 and Beclin1 increased gradually after 2 days p.i. and began to decrease at 5 days after infection, respectively.

CONCLUSION

These results suggested that HCMV infection can facilitate the autophagy and autophagy level may decrease in latent phase. More studies on the relationship between HCMV latency and autophagy are needed to determine the role of autophagy in HCMV latent infection that may help find out a therapeutic approach for clinical treatment.

摘要

背景

人巨细胞病毒(HCMV)感染非常常见,且潜伏感染日后可能会重新激活。与其他疱疹病毒类似,它可改变细胞内环境以利于病毒复制和存活。本研究旨在探讨HCMV感染对人急性单核细胞白血病细胞系(THP - 1细胞)自噬的影响。

方法

采用逆转录聚合酶链反应(RT - PCR)、蛋白质印迹法及透射电子显微镜(TEM)检测自噬水平。将自噬相关蛋白Beclin 1、Atg5及轻链3(LC3)的浓度与肌动蛋白水平进行比较计数。

结果

与对照THP - 1细胞相比,感染后1至5天(d),Beclin1、Atg5及LC3II mRNA的表达水平逐渐升高,随后逐渐下降。然而,蛋白质印迹分析结果显示,感染后1天LC3II水平逐渐升高,感染后7天下降。但Atg5和Beclin1水平分别在感染后2天逐渐升高,感染后5天开始下降。

结论

这些结果表明,HCMV感染可促进自噬,且自噬水平在潜伏期可能降低。需要对HCMV潜伏与自噬之间的关系进行更多研究,以确定自噬在HCMV潜伏感染中的作用,这可能有助于找到临床治疗的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86f5/5682777/09b2d52b4c8b/medi-96-e8298-g002.jpg

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