WRN 通过 ATM/p53 信号通路抑制氧化应激诱导的人晶状体上皮细胞凋亡，其表达受 DNA 甲基化下调。

WRN inhibits oxidative stress-induced apoptosis of human lensepithelial cells through ATM/p53 signaling pathway and its expression is downregulated by DNA methylation.

机构信息

Ophthalmology Department, The First Affiliated Hospital of Jinan University Guangzhou, No.601 Huangpu Avenue West, Guangzhou, Guangdong Province, China.

Ophthalmology Department, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui Province, China.

出版信息

Mol Med. 2020 Jul 8;26(1):68. doi: 10.1186/s10020-020-00187-x.

DOI:10.1186/s10020-020-00187-x

PMID:32640976

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7341633/

Abstract

BACKGROUND

Apoptosis and oxidative stress are the main etiology of age related cataract (ARC). This article aims to investigate the role of WRN in lens epithelial cells (LECs).

METHODS

We estimated the methylation level of WRN in anterior lens capsule tissues of ARC patients. SRA01/04 (LECs) cells were treated with HO or combined with 5-aza-2-deoxycytidine (5-Aza-CdR) or chloroquine. CCK8 and flow cytometry were performed to explore proliferation and apoptosis. The content of ROS was detected by fluorescent probe DCFH-DA. The gene and protein expression was assessed by quantitative real-time PCR or western blot.

RESULTS

WRN was down-regulated and the methylation level of WRN was increased in the anterior lens capsule tissues. WRN overexpression and 5-Aza-CdR enhanced proliferation and repressed apoptosis and oxidative stress of SRA01/04 cells. 5-Aza-CdR enhanced WRN expression. WRN knockdown inhibited proliferation and promoted apoptosis and oxidative stress of SRA01/04 cells, which was rescued by 5-Aza-CdR. WRN overexpression and 5-Aza-CdR repressed ATM/p53 signaling pathway. Furthermore, chloroquine inhibited proliferation and promoted apoptosis and oxidative stress of SRA01/04 cells by activating ATM/p53 signaling pathway. The influence conferred by chloroquine was abolished by WRN overexpression.

CONCLUSION

Our study reveals that DNA methylation mediated WRN inhibits apoptosis and oxidative stress of human LECs through ATM/p53 signaling pathway.

摘要

背景

细胞凋亡和氧化应激是年龄相关性白内障（ARC）的主要病因。本文旨在研究 WRN 在晶状体上皮细胞（LECs）中的作用。

方法

我们评估了 ARC 患者前晶状体囊组织中 WRN 的甲基化水平。SRA01/04（LECs）细胞用 HO 或与 5-氮杂-2′-脱氧胞苷（5-Aza-CdR）或氯喹联合处理。通过 CCK8 和流式细胞术研究增殖和凋亡。通过荧光探针 DCFH-DA 检测 ROS 含量。通过定量实时 PCR 或 Western blot 评估基因和蛋白表达。

结果

WRN 下调，前晶状体囊组织中 WRN 的甲基化水平增加。WRN 过表达和 5-Aza-CdR 增强了 SRA01/04 细胞的增殖，抑制了凋亡和氧化应激。5-Aza-CdR 增强了 WRN 的表达。WRN 敲低抑制了 SRA01/04 细胞的增殖，促进了凋亡和氧化应激，5-Aza-CdR 挽救了这一作用。WRN 过表达和 5-Aza-CdR 抑制了 ATM/p53 信号通路。此外，氯喹通过激活 ATM/p53 信号通路抑制了 SRA01/04 细胞的增殖，促进了凋亡和氧化应激。WRN 过表达消除了氯喹的影响。