Wei Youhua, Wang Zhijun, Wei Li, Li Shen, Qiu Xuemei, Liu Chengwen
Department of Medical Heredity and Prenatal Screening, Zaozhuang Maternal and Child Health Care Hospital, Zaozhuang, Shandong 277100, P.R. China.
Department of Obstetrics and Gynecology, Zaozhuang Maternal and Child Health Care Hospital, Zaozhuang, Shandong 277100, P.R. China.
Exp Ther Med. 2021 Apr;21(4):359. doi: 10.3892/etm.2021.9790. Epub 2021 Feb 13.
MicroRNA (miR)-874-3p is a newly identified miRNA that is involved in several pathological processes, including cancer, myocardial infarction, bone formation and erectile dysfunction. However, the role of miR-874-3p in polycystic ovary syndrome (PCOS) and granulosa cell (GC) apoptosis is not completely understood. The present study investigated the expression profile of miR-874-3p in PCOS by reverse transcription- quantitative PCR and the GC apoptosis by flow cytometry analysis. miR-874-3p expression was significantly upregulated in GCs isolated from patients with PCOS compared with patients without PCOS. In addition, miR-874-3p expression was positively correlated with GC apoptosis and testosterone levels in both patients with PCOS and patients without PCOS. Therefore, the present study also aimed to investigate the effects of miR-874-3p on testosterone-induced GC apoptosis. Compared with vehicle-treated GCs, miR-874-3p expression levels were significantly increased in testosterone-treated GCs, which was inhibited by the androgen receptor antagonist flutamide. GCs were transfected with either the miR-874-3p mimic or a miR-874-3p inhibitor. Compared with the control group, miR-874-3p mimic significantly enhanced GC apoptosis, whereas miR-874-3p inhibitor significantly decreased GC apoptosis. Moreover, histone deacetylase (HDAC) activity and HDAC1 expression levels were decreased in testosterone-treated GCs compared with vehicle-treated GCs. HDAC1 overexpression significantly attenuated the proapoptotic effects of testosterone. Additionally, miR-874-3p mimic and inhibitor significantly decreased and increased HDAC1 expression levels, respectively, compared with the control group. miR-874-3p inhibitor failed to attenuate HDAC1 overexpression-induced GC apoptosis. Furthermore, compared with the control group, testosterone treatment notably increased p53 expression and acetylation. Compared with the control group, western blotting analysis showed that miR-874-3p mimic notably increased p53 expression and acetylation, whereas miR-874-3p inhibitor markedly decreased p53 expression and acetylation. However, miR-874-3p inhibitor did not further decrease p53 acetylation and expression in cell overexpressing HDAC1. Collectively, the results of the present study indicated that miR-874-3p was upregulated in PCOS and promoted testosterone-induced GC apoptosis by suppressing HDAC1-mediated p53 deacetylation. Therefore, the present study improved the current understanding of the pathogenesis of PCOS and GC apoptosis.
微小RNA（miR）-874-3p是一种新发现的微小RNA，参与多种病理过程，包括癌症、心肌梗死、骨形成和勃起功能障碍。然而，miR-874-3p在多囊卵巢综合征（PCOS）和颗粒细胞（GC）凋亡中的作用尚未完全明确。本研究通过逆转录定量PCR检测PCOS中miR-874-3p的表达谱，并通过流式细胞术分析GC凋亡情况。与非PCOS患者相比，PCOS患者分离的GC中miR-874-3p表达显著上调。此外，PCOS患者和非PCOS患者中，miR-874-3p表达均与GC凋亡和睾酮水平呈正相关。因此，本研究还旨在探讨miR-874-3p对睾酮诱导的GC凋亡的影响。与溶剂处理的GC相比，睾酮处理的GC中miR-874-3p表达水平显著升高，雄激素受体拮抗剂氟他胺可抑制该升高。将GC分别转染miR-874-3p模拟物或miR-874-3p抑制剂。与对照组相比，miR-874-3p模拟物显著增强GC凋亡；而miR-874-3p抑制剂显著降低GC凋亡。此外，与溶剂处理的GC相比，睾酮处理的GC中组蛋白去乙酰化酶（HDAC）活性和HDAC1表达水平降低。HDAC1过表达显著减弱睾酮的促凋亡作用。此外，与对照组相比，miR-874-3p模拟物和抑制剂分别显著降低和升高HDAC1表达水平。miR-874-3p抑制剂未能减弱HDAC1过表达诱导的GC凋亡。此外，与对照组相比，睾酮处理显著增加p53表达和乙酰化。与对照组相比，蛋白质印迹分析显示，miR-874-3p模拟物显著增加p53表达和乙酰化；而miR-874-3p抑制剂显著降低p53表达和乙酰化。然而，miR-874-3p抑制剂未进一步降低HDAC1过表达细胞中p53的乙酰化和表达。总体而言，本研究结果表明，PCOS中miR-874-3p上调，并通过抑制HDAC1介导的p53去乙酰化促进睾酮诱导的GC凋亡。因此，本研究增进了对PCOS发病机制和GC凋亡的当前认识。
