CircPAG1 Inhibits the High Glucose-Induced Lens Epithelial Cell Injury by Sponging miR-630 and Upregulating EPHA2

ABSTRACT

Background: Circular RNAs (circRNAs) have been considered as vital regulators in the progression of human ocular diseases, including diabetic cataract (DC). This report was designed to research the biological role of circRNA phosphoprotein associated with glycosphingolipid-enriched microdomains 1 (circPAG1) in high glucose (HG)-induced lens epithelial damages.

Methods: Lens epithelial damage in DC was investigated by the effects of 25 mM glucose (HG) on human lens epithelial cells (HLE-B3). CircPAG1, microRNA-630 (miR-630), and ephrin type-A receptor 2 (EPHA2) levels were examined by the quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation analysis was performed by 3-(4, 5-dimethylthiazol-2-y1)-2, 5-diphenyl tetrazolium bromide (MTT) assay and colony formation assay. Cell apoptosis was measured through flow cytometry. Protein levels were detected using western blot. Oxidative stress was determined by malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) levels via the corresponding kits. Dual-luciferase reporter and RNA immunoprecipitation (RIP) and RNA pull-down assays were used for target binding analysis.

Results: CircPAG1 expression was downregulated in lens samples of DC patients and HG-treated lens epithelial cells. HG inhibited cell growth but promoted apoptosis and oxidative stress in HLE-B3 cells, while circPAG1 overexpression relieved these damages. Moreover, circPAG1 was identified as a molecular sponge for miR-630. HG-induced cell injury was also attenuated by the inhibition of miR-630, and the function of circPAG1 was related to its sponge effect on miR-630. In addition, miR-630 directly targeted EPHA2 and circPAG1 could regulate the EPHA2 expression via sponging miR-630. Furthermore, we found that the protective role of circPAG1 against the HG-induced cell injury was ascribed to the upregulation of EPHA2.

Conclusion: Our evidence suggested that circPAG1 alleviated cell damages in HG-treated human lens epithelial cells by regulating the miR-630/EPHA2 axis.

KEYWORDS:

• CircPAG1
• miR-630
• EPHA2
• diabetic cataract

Ethics approval and consent of participants

Written informed consent was obtained from patients with approval by the Institutional Review Board in Enze Medical Center.