MiR-34a-5p Negatively Regulates Oxidative Stress on Lens Epithelial Cells by Silencing GPX3 – A Novel Target
Year: 2022
Type: dataset
Abstract: 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. The three candidate miRNAs were screened by CCK-8 assays after transfection of mimics or inhibitor in H2O2-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. The viability of H2O2-treated HLE-B3 cells were weakened by up-regulated miR-34a-5p. Cell apoptosis and oxidative damage were also induced by overexpression of miR-34a-5p. GPX3 and SRC were identified as target genes of miR-34a-5p by combined analysis of proteomic and bioinformatics, while GPX3 was selected for further research for its connection with anti-oxidation. Western blot and qRT-PCR tests proved that GPX3 is negatively regulated by miR-34a-5p. Dual-luciferase reporter assay verified that GPX3 is the direct target of miR-34a-5p. The increased oxidative stress induced by transfection of miR-34a-5p mimics in H2O2-treated HLE-B3 cells was attenuated by overexpression of GPX3. 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.
Source: Figshare
Authors: Wang, Song, Yu, Mengsi, Yan, Hong, Liu, Jun, Guo, Chenjun
Institutions: University of South Carolina, Shenyang University of Technology, Tianjin University, First Affiliated Hospital Zhejiang University, Wuhan University, Xi'an Jiaotong University, Guangzhou University of Chinese Medicine, Shandong University, University of California, San Francisco, Capital Medical University, Xiamen University, Cornell University, Sichuan University, Academy of Military Medical Sciences, Tongren Hospital, Suzhou Municipal Hospital, Shandong Provincial Hospital, Indiana University Bloomington, Mayo Clinic Health System, First Affiliated Hospital of GuangXi Medical University, Peking University First Hospital, Chinese Academy of Agricultural Sciences, Chongqing Population and Family Planning Science and Technology Research Institute, Second Xiangya Hospital of Central South University, Huazhong University of Science and Technology, China Agricultural University, Zhejiang University, Yangzhou University
Language: English
Subfield: Cancer Research
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W4394496648