ROS/p53/miR‑335‑5p/Sp1 axis modulates the migration and epithelial to mesenchymal transition of JEG‑3 cells

Lu,Wei; Ma,Yu‑Yan; Shao,Qian‑Qian; Liang,Jie; Qi,Tong‑Tong; Huang,Yi; Wang,Qing‑Jie

doi:10.3892/mmr.2019.10901

ROS/p53/miR‑335‑5p/Sp1 axis modulates the migration and epithelial to mesenchymal transition of JEG‑3 cells

Authors:
- Wei Lu
- Yu‑Yan Ma
- Qian‑Qian Shao
- Jie Liang
- Tong‑Tong Qi
- Yi Huang
- Qing‑Jie Wang
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Affiliations: Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, P.R. China, The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Institute of Basic Medical Sciences, Qilu Hospital of Shandong University, Jinan, Shandong 250012, P.R. China, Central Sterile Supply Department, People's Hospital of Fangzi, Weifang, Shandong 261200, P.R. China, School of Pharmaceutical Science, Shandong University, Jinan, Shandong 250012, P.R. China
Published online on: December 20, 2019 https://doi.org/10.3892/mmr.2019.10901
Pages: 1208-1216
Copyright: © Lu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Differential expression of microRNA (miR)‑335‑5p, a key tumor suppressor, has been detected in pre‑eclampsia (PE) placentas. However, the role of miR‑335‑5p in the pathogenesis of PE and the factor modulating its aberrant expression remain unknown. The present study used JEG‑3 cells in vitro to investigate these mechanisms. The role of miR‑335‑5p in proliferation, apoptosis and migration of JEG‑3 cells was investigated using MTT, Annexin V‑FITC/PI, Transwell migration and wound healing assays, respectively. miR‑335‑5p expression levels were analyzed using reverse transcription‑quantitative PCR. The expression levels of E‑cadherin, N‑cadherin, Snail, specificity protein 1 (Sp1) and p53 were assessed using western blot analysis. Cell viability analysis was performed using the Cell Counting Kit‑8 assay. The intracellular reactive oxygen species (ROS) levels were detected using a 2,7‑dichlorodihydrofluorescein diacetate assay. The present results suggested that miR‑335‑5p did not affect the proliferation or apoptotic rate of JEG‑3 cells. Overexpression of miR‑335‑5p significantly inhibited the migration of JEG‑3 cells, decreased the expression levels of Sp1, N‑cadherin and Snail, and increased E‑cadherin expression. Sp1 silencing produced similar results in JEG‑3 cells. H2O2 significantly increased the intracellular ROS levels and miR‑335‑5p expression, whereas N‑acetyl‑cysteine pretreatment prior to H2O2 treatment reversed the increases in miR‑335‑5p expression. Knockdown of p53 significantly decreased the expression levels of miR‑335‑5p in JEG‑3 cells and in H2O2‑treated cells. The present results suggested that miR‑335‑5p expression levels in trophoblast cells could be increased by ROS in a p53‑dependent manner, leading to the downregulation of Sp1 and subsequent inhibition of epithelial to mesenchymal transition and cell migration. The present results may provide novel evidence on the etiology of PE.

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