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

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

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 (H2O2). 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 H2O2-treated SRA01/04 cells. H2O2 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 H2O2-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 H2O2-treated SRA01/04 cells.

Conclusion: The increased expression of circSTK39 ameliorated H2O2-induced SRA01/04 cell injury through the miR-125a-5p/ERCC6 pathway.

Keywords: Age-related cataract; ERCC6; circSTK39; miR-125a-5p.

MeSH terms

  • Apoptosis / genetics
  • Cataract* / genetics
  • Cataract* / metabolism
  • Cell Proliferation
  • DNA Helicases
  • DNA Repair Enzymes
  • Epithelial Cells / metabolism
  • Humans
  • Hydrogen Peroxide / metabolism
  • Hydrogen Peroxide / toxicity
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Oxidative Stress
  • Poly-ADP-Ribose Binding Proteins
  • RNA, Circular / genetics
  • RNA, Circular / metabolism

Substances

  • Hydrogen Peroxide
  • MicroRNAs
  • RNA, Circular
  • ERCC6 protein, human
  • Poly-ADP-Ribose Binding Proteins
  • DNA Helicases
  • DNA Repair Enzymes