Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

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

Jing Wang¹, Yanping Zhao²

¹Department of Gynecology and Obstetrics, Chengdu Second Peopleâ€™s Hospital, Chengdu, Sichuan Province 610017, China; ²Department of Obstetrics, Hubei Provincial Hospital of Integrated Chinese &Western Medicine, Wuhan, Hubei Province 430000, China.

For correspondence:- Yanping Zhao Email: yp_zhao88@163.com Tel:+862765600775

Accepted: 29 January 2023 Published: 28 February 2023

Citation: Wang J, Zhao Y. Knockdown of PRUNE2 alleviates hypoxia-induced oxidative stress inhibits cell proliferation in trophoblast cells, and reverses LY294002-induced PI3K/AKT pathway inhibition. Trop J Pharm Res 2023; 22(2):305-311 doi: 10.4314/tjpr.v22i2.12

© 2023 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

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 μM 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.

Keywords: Preeclampsia, PRUNE2, PI3K/AKT pathway, Trophoblast cells, Oxidative stress