AIM:To study microRNAs(miRNAs)and their potential effects in high glucose-induced human retinal pigment epithelial cell damage.METHODS:We screened the GSE52233 miRNA expression dataset for differentially expressed miR...AIM:To study microRNAs(miRNAs)and their potential effects in high glucose-induced human retinal pigment epithelial cell damage.METHODS:We screened the GSE52233 miRNA expression dataset for differentially expressed miRNAs(DEMs).The target genes of the top 10 DEMs were predicted using miRWalk 2.0 database,followed by function enrichment and protein-protein interaction analysis.miRNA expression was determined in the human retinal pigment epithelial cell line ARPE-19 treated with high glucose(HG)by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).Cell proliferation was determined using cell counting kit(CCK)-8 assay.Cell cycle,apoptosis,and reactive oxygen species(ROS)levels were determined by flow cytometry.The direct interaction between miRNA and targets was validated using dual-luciferase reporter assay.RESULTS:Thirty-nine DEMs were screened,and we predicted 125 miRNA-mRNA pairs for the top 10 DEMs,including 119 target genes of seven DEMs such as miR-346,which was upregulated in diabetic retinopathy(DR).miR-346 target genes were substantially enriched in the regulation of intracellular transport and retinoic acidinducible gene I(RIG-I)-like receptor signaling pathway.Expression of three upregulated and downregulated miRNAs were verified by qRT-PCR in HG-treated ARPE-19 cells.Expression of miR-346 was elevated in HG treated ARPE-19 cells in a dose-dependent manner.HG inhibited cell proliferation and induced apoptosis,which were partly reversed by transfecting an miR-346 inhibitor,which even decreased the ROS levels elevated due to HG.Argonaute 2(AGO2)was a target of miR-346.CONCLUSION:miR-346 is a key miRNA and plays an important role in HG-induced damage in human retinal pigment epithelial cells.展开更多
基金Supported by the Social Development Project of Shaanxi Provincial Department of Science and Technology(No.2020SF-167)Supporting Fund Project of Shaanxi Provincial Department of Science and Technology Agency Project(No.2022SF-502)Xi’an Medical University 2022 Annual Scientific Research Capacity Improvement Plan Project(No.2022NLTS104).
文摘AIM:To study microRNAs(miRNAs)and their potential effects in high glucose-induced human retinal pigment epithelial cell damage.METHODS:We screened the GSE52233 miRNA expression dataset for differentially expressed miRNAs(DEMs).The target genes of the top 10 DEMs were predicted using miRWalk 2.0 database,followed by function enrichment and protein-protein interaction analysis.miRNA expression was determined in the human retinal pigment epithelial cell line ARPE-19 treated with high glucose(HG)by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).Cell proliferation was determined using cell counting kit(CCK)-8 assay.Cell cycle,apoptosis,and reactive oxygen species(ROS)levels were determined by flow cytometry.The direct interaction between miRNA and targets was validated using dual-luciferase reporter assay.RESULTS:Thirty-nine DEMs were screened,and we predicted 125 miRNA-mRNA pairs for the top 10 DEMs,including 119 target genes of seven DEMs such as miR-346,which was upregulated in diabetic retinopathy(DR).miR-346 target genes were substantially enriched in the regulation of intracellular transport and retinoic acidinducible gene I(RIG-I)-like receptor signaling pathway.Expression of three upregulated and downregulated miRNAs were verified by qRT-PCR in HG-treated ARPE-19 cells.Expression of miR-346 was elevated in HG treated ARPE-19 cells in a dose-dependent manner.HG inhibited cell proliferation and induced apoptosis,which were partly reversed by transfecting an miR-346 inhibitor,which even decreased the ROS levels elevated due to HG.Argonaute 2(AGO2)was a target of miR-346.CONCLUSION:miR-346 is a key miRNA and plays an important role in HG-induced damage in human retinal pigment epithelial cells.
