Knockdown of PRUNE2 alleviates hypoxia-induced oxidative stress inhibits cell proliferation in trophoblast cells, and reverses LY294002-induced PI3K/AKT pathway inhibition

Purpose: To investigate the probable effect of PRUNE2 on viability, motility, and oxidative stress of hypoxia-induced trophoblast cells, and the mechanism of action involved. Methods: Trophoblast cells were treated with 250 mu M CoCl2 for 24 h to simulate hypoxia. Then, a cell counting kit (CCK-8), colony formation, and flow cytometry (FCM) were used to determine the role of PRUNE2 in cell viability. Wound closure, as well as Transwell-migration assay, were conducted to evaluate cell motility. Superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), and myeloperoxidase (MPO) levels were determined using the appropriate kits, while the effects of PRUNE2 on PI3K/AKT pathway in trophoblast cells were assessed by immunoblot assay. LY294002 was used to inhibit PI3K/AKT pathway in trophoblast cells. Results: The hypoxia-induced trophoblast cell model was successfully constructed. The data showed that PRUNE2 was overexpressed in hypoxia-induced trophoblast cells, and that ablation of PRUNE2 stimulated proliferation as well as motility of hypoxia-induced trophoblast cells (p < 0.01). Depletion of PRUNE2 reduced oxidative stress in the cells. PRUNE2 mediated PI3K/AKT pathway and thus affected the proliferation, motility, and oxidative stress of trophoblast cells (p < 0.01). In addition, depletion of PRUNE2 reversed PI3K/AKT signaling and the inhibition of trophoblast proliferation induced by t PI3K/AKT inhibitor LY294002. Conclusion: PRUNE2 depletion alleviates hypoxia-induced oxidative stress, inhibits cell proliferation in trophoblast cells and reverses LY294002-induced PI3K/AKT pathway inhibition. The findings suggest that PRUNE2 could act as a target treatment for PE.