Microarray Analysis of Small Extracellular Vesicle-Derived miRNAs Involved in Oxidative Stress of RPE Cells.

Oxidative Medicine and Cellular Longevity
Ke Mao, Xingwei Wu

Abstract

The aim of this study was to investigate the miRNA profiles of nanosized small extracellular vesicles (sEVs) from human retinal pigment epithelial (RPE) cells under oxidative damage. ARPE-19 cells were cultured with ox-LDL (100 mg/L) or serum-free medium for 48 hours, sEVs were then extracted, and miRNA sequencing was conducted to identify the differentially expressed genes (DEGs) between the 2 groups. RNA sequence results were validated using quantitative real-time PCR. The Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes pathway, and ingenuity pathway analyses (IPA) were performed for the DEGs. Results revealed that oxidative stress inhibited RPE cell viability and promoted sEV secretion. A total of 877 DEGs from sEVs were identified, of which 272 were downregulated and 605 were upregulated. In total, 66 enriched GO terms showed that the 3 most significant enrichment terms were cellular processes (biological processes), cell (cellular component), and catalytic activity (molecular function). IPA were used to explore DEGs associated with oxidation damage and further construct a miRNA-target regulatory network. This study identified several DEGs from oxidation-stimulated RPE cells, which may act as potentia...Continue Reading

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Methods Mentioned

BETA
Assay
PCR
Transmission

Software Mentioned

TargetScan
SPSS
Bowtie2
limma
miRanda
ImageJ

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