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MiR-34a-5p Negatively Regulates Oxidative Stress on Lens Epithelial Cells by Silencing GPX3 – A Novel Target

Song Wang, Mengsi Yu, Hong Yan, Jun Liu, Chenjun Guo

2022Current Eye Research12 citationsDOIOpen Access PDF

Abstract

PURPOSE: MiR-34a-5p is reported to be related with age-related nuclear cataract. This study investigated the mechanism of miR-34a-5p in the regulation of oxidative stress on lens epithelial cells. METHODS: -treated HLE-B3 cells. The apoptosis, ROS level and GPX activity of HLE-B3 cells transfected with miR-34a-5p mimics or inhibitor were analysed by flow cytometry, cellular ROS and GPX activity test. The target genes of miR-34a-5p were predicted by proteomic and bioinformatic analysis. The relationship between miR-34a-5p and GPX3 were internally validated by qRT-PCR and Western blot and externally verified by dual-luciferase reporter assay. The effect of miR-34a-5p-GPX3 axis on regulation of oxidative stress in HLE-B3 cells were conducted by overexpression of GPX3 and tested by flow cytometry analysis, cellular ROS and GPX detection. RESULTS: -treated HLE-B3 cells was attenuated by overexpression of GPX3. CONCLUSIONS: MiR-34a-5p is a negative regulator of oxidative stress on lens epithelial cells and the mechanism is by silencing the expression of GPX3. These data suggest that miR-34a-5p may be a potential novel therapeutic target for the prevention and treatment of age-related cataract.

Topics & Concepts

Oxidative stressTransfectionWestern blotFlow cytometryMolecular biologyApoptosisCell biologyGene silencingChemistryBiologyCell cultureBiochemistryGeneGeneticsConnexins and lens biologyIntraocular Surgery and LensesAldose Reductase and Taurine
MiR-34a-5p Negatively Regulates Oxidative Stress on Lens Epithelial Cells by Silencing GPX3 – A Novel Target | Litcius