Li Jun, Wu Yan, Liu Hui
Fetal Heart Monitoring Unit, Laiwu Maternal and Child Health Hospital, Laiwu, Shandong 271100, P.R. China.
The Fifth Department of Obstetrics and Gynecology, Laiwu Maternal and Child Health Hospital, Laiwu, Shandong 271100, P.R. China.
Exp Ther Med. 2020 Jul;20(1):418-426. doi: 10.3892/etm.2020.8719. Epub 2020 May 6.
The present study aimed to investigate the role of miR-338-3p in pregnancy-induced hypertension (PIH), and its effects on human trophoblast cells . Quantitative real-time PCR was used to detect miR-338-3p expression. Human trophoblast HTR8/SVneo cells were transfected with miR-338-3p mimics. Effects of miR-338-3p on cell proliferation, invasion and metastasis, and anoikis resistance were detected by CCK-8 assay, Transwell chamber assay, flow cytometry and western blot analysis, respectively. Bioinformatics analysis was performed to predict the target of miR-338-3p, and the results were confirmed by dual luciferase reporter assay. The expression level of miR-338-3p was significantly upregulated in the peripheral blood and placenta of PIH patients. CCK-8 assay showed that miR-338-3p mimics inhibited the proliferation of HTR8/SVneo cells at indicated time points. Flow cytometry showed that miR-338-3p transfection significantly increased the Ki-67 expression in the HTR8/SVneo cells, indicating enhanced cell proliferation. Transwell chamber assay and western blot analysis showed that the invasion and metastatic abilities of the HTR8/SVneo cells were significantly decreased in the miR-338-3p transfection group, as well as expression levels of MMP-2 and MMP-9. Bioinformatics analysis and dual luciferase reporter assay indicated that is a target gene of miR-338-3p. Our results suggest that miR-338-3p is significantly increased in the peripheral blood and placenta of PIH patients, which is correlated with the disease development. miR-338-3p inhibits proliferation, invasion and metastasis, and apoptosis resistance of human trophoblast cells by targeting .
本研究旨在探讨miR-338-3p在妊娠期高血压疾病(PIH)中的作用及其对人滋养层细胞的影响。采用定量实时PCR检测miR-338-3p表达。用miR-338-3p模拟物转染人滋养层HTR8/SVneo细胞。分别通过CCK-8法、Transwell小室法、流式细胞术和蛋白质免疫印迹分析检测miR-338-3p对细胞增殖、侵袭转移及失巢凋亡抗性的影响。进行生物信息学分析以预测miR-338-3p的靶标,并通过双荧光素酶报告基因检测进行验证。PIH患者外周血和胎盘中miR-338-3p的表达水平显著上调。CCK-8法显示,miR-338-3p模拟物在指定时间点抑制HTR8/SVneo细胞的增殖。流式细胞术显示,miR-338-3p转染显著增加HTR8/SVneo细胞中Ki-67的表达,表明细胞增殖增强。Transwell小室法和蛋白质免疫印迹分析显示,miR-338-3p转染组HTR8/SVneo细胞的侵袭和转移能力显著降低,同时MMP-2和MMP-9的表达水平也降低。生物信息学分析和双荧光素酶报告基因检测表明, 是miR-338-3p的靶基因。我们的结果表明,PIH患者外周血和胎盘中miR-338-3p显著增加,这与疾病发展相关。miR-338-3p通过靶向 抑制人滋养层细胞的增殖、侵袭转移及凋亡抗性。