异位 circSTK39 表达通过 miR-125a-5p/ERCC6 通路减轻过氧化氢诱导的人晶状体上皮细胞凋亡和氧化应激。

Ectopic circSTK39 Expression Ameliorates Hydrogen Peroxide-Induced Human Lens Epithelial Cell Apoptosis and Oxidative Stress through the miR-125a-5p/ERCC6 Pathway.

机构信息

Department of Ophthalmology, Jingzhou Hospital, Yangtze University (Jingzhou Central Hospital), Jingzhou, Hubei, China.

Department of Pathology, School of Medicine, Yangtze University, Jingzhou, Hubei, China.

出版信息

Curr Eye Res. 2023 Mar;48(3):278-288. doi: 10.1080/02713683.2022.2143529. Epub 2022 Nov 15.

DOI:10.1080/02713683.2022.2143529

PMID:36322706

Abstract

PURPOSE

More and more studies suggest that circular RNA (circRNA) is involved in the pathogenesis of age-related cataract (ARC). CircSTK39, a circular RNA, has inhibitory effects on cancer progression. However, there is no data regarding the role of circSTK39 in ARC occurrence and the underlying mechanism.

METHODS

ARC cell model was established by inducing lens epithelial cells (SRA01/04) using hydrogen peroxide (HO). CircSTK39, microRNA-125a-5p (miR-125a-5p), and ERCC excision repair 6, chromatin remodeling factor (ERCC6) expression were detected by quantitative real-time polymerase chain reaction. Western blot was conducted to assess protein expression. Cell viability, proliferation, and apoptosis were investigated by cell counting kit-8 assay, 5-Ethynyl-29-deoxyuridine assay, and flow cytometry analysis, respectively. Oxidative stress was evaluated using commercial kits. Dual-luciferase reporter assay, RNA immunoprecipitation assay, and RNA pull-down assay were used to identify the relationship between miR-125a-5p and circSTK39 or ERCC6.

RESULTS

CircSTK39 and ERCC6 expression were significantly downregulated, but miR-125a-5p expression was upregulated in the lens tissues of ARC patients and HO-treated SRA01/04 cells. HO treatment led to decreased cell proliferation and increased cell apoptosis and oxidative stress, accompanied by the increases of C-caspase3 and Bax expression and the decrease of Bcl-2 expression; however, these effects were reversed after circSTK39 overexpression. MiR-125a-5p was found to participate in HO-triggered cell damage by interacting with circSTK39. Additionally, ERCC6 silencing inhibited circSTK39 overexpression-mediated action. Importantly, circSTK39 regulated ERCC6 expression by interaction with miR-125a-5p in HO-treated SRA01/04 cells.

CONCLUSION

The increased expression of circSTK39 ameliorated HO-induced SRA01/04 cell injury through the miR-125a-5p/ERCC6 pathway.

摘要

目的

越来越多的研究表明，环状 RNA（circRNA）参与年龄相关性白内障（ARC）的发病机制。环状 STK39 是一种环状 RNA，对癌症进展具有抑制作用。然而，关于 circSTK39 在 ARC 发生中的作用及其潜在机制尚无数据。

方法

用双氧水（HO）诱导晶状体上皮细胞（SRA01/04）建立 ARC 细胞模型。通过实时定量聚合酶链反应检测 circSTK39、微小 RNA-125a-5p（miR-125a-5p）和 ERCC 切除修复 6、染色质重塑因子（ERCC6）的表达。通过 Western blot 检测蛋白表达。通过细胞计数试剂盒-8 测定法、5-乙炔基-29-脱氧尿苷测定法和流式细胞术分析分别评估细胞活力、增殖和凋亡。使用商业试剂盒评估氧化应激。双荧光素酶报告基因测定、RNA 免疫沉淀测定和 RNA 下拉测定用于鉴定 miR-125a-5p 与 circSTK39 或 ERCC6 之间的关系。

结果

在 ARC 患者的晶状体组织和 HO 处理的 SRA01/04 细胞中，circSTK39 和 ERCC6 的表达明显下调，而 miR-125a-5p 的表达上调。HO 处理导致细胞增殖减少，细胞凋亡和氧化应激增加，同时 C-caspase3 和 Bax 表达增加，Bcl-2 表达减少；然而，circSTK39 过表达后这些作用得到逆转。发现 miR-125a-5p 通过与 circSTK39 相互作用参与 HO 触发的细胞损伤。此外，沉默 ERCC6 抑制了 circSTK39 过表达介导的作用。重要的是，circSTK39 通过与 HO 处理的 SRA01/04 细胞中的 miR-125a-5p 相互作用调节 ERCC6 的表达。