miR-208a-3p promotes hypoxia-induced oxidative stress and cellular dysfunction by targeting ETS1 in trophoblast and endothelial cells

This study investigated the role of miR-208a-3p in hypoxia-induced trophoblast and endothelial dysfunction in an in vitro model of GH. HTR-8/SVneo and HUVECs were exposed to hypoxia (1% O₂, 24 h) to establish a GH cell model. miR-208a-3p inhibitor and si-ETS1 were transfected into cells. qPCR, CCK-8, Transwell, dual-luciferase reporter assay, and ferroptosis-related indicator detection were performed. Bioinformatics analysis using TargetScan, miRDB, and miRWalk databases predicted downstream target genes, followed by GO and KEGG enrichment analyses. Hypoxia significantly upregulated miR-208a-3p expression and decreased the GSH/GSSG ratio, while increasing Fe 2 ⁺ and MDA levels and decreasing GPX4 and SLC7A11 expression. miR-208a-3p inhibitor reversed these ferroptosis-related changes. Additionally, miR-208a-3p inhibitor partially restored the hypoxia-induced reduction of sFlt-1 and reversed the hypoxia-induced upregulation of Hsp90, while also recovering invasion and proliferation in HTR-8/SVneo cells and migration and proliferation in HUVECs. Bioinformatics analysis identified ETS1 as a direct target of miR-208a-3p, confirmed by dual-luciferase assay. ETS1 silencing induced similar ferroptosis-related changes and functional impairments. In this in vitro model, miR-208a-3p promotes hypoxia-induced ferroptosis-related changes and impairs trophoblast and endothelial cell function by targeting ETS1, suggesting a potential mechanism relevant to GH that warrants clinical validation.