Wang Mingming, Sun Jing, Xu Bo, Chrusciel Marcin, Gao Jun, Bazert Maciej, Stelmaszewska Joanna, Xu Yunyun, Zhang Hongwen, Pawelczyk Leszek, Sun Fei, Tsang Suk Ying, Rahman Nafis, Wolczynski Slawomir, Li Xiangdong
Beijing Advanced Innovation Center for Food Nutrition and Human Health, China Agricultural University, Beijing, China.
State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
Endocrinology. 2018 Jan 1;159(1):297-309. doi: 10.1210/en.2017-00219.
The goal of this study was to characterize the function of microRNA-27a-3p (miR-27a-3p) in polycystic ovary syndrome (PCOS). miR-27a-3p expression was analyzed in excised granulosa cells (GCs) from 21 patients with PCOS and 12 normal patients undergoing in vitro fertilization cycle treatments and in 17 nontreated cuneiform ovarian resection PCOS samples and 13 control ovarian samples from patients without PCOS. We found that the expression of miR-27a-3p was significantly increased in both excised GCs and the ovaries of patients with PCOS compared with the controls. Insulin treatment of the human granulosa-like tumor cell line (KGN) resulted in decreased downregulated expression of miR-27a-3p, and this effect appeared to be mediated by signal transducer and activator of transcription STAT1 and STAT3. The overexpression of miR-27a-3p in KGN cells inhibited SMAD5, which in turn decreased cell proliferation and promoted cell apoptosis. After KGN cells were stimulated with insulin for 48 hours, there was increased expression of SMAD5 protein and decreased apoptosis. Additionally, knockdown/overexpression of SMAD5 in KGN cells reduced/increased cell number and promoted/inhibited cell apoptosis. Insulin-stimulated primary GCs isolated from patients with PCOS, in contrast to normal GCs or KGN cells, did not exhibit decreased miR-27a-3p expression. The differences in the expression levels in KGN cells and human PCOS GCs are likely explained by increased miR-27a-3p expression in the GCs caused by insulin resistance in PCOS. Taken together, our data provided evidence for a functional role of miR-27a-3p in the GCs' dysfunction that occurs in patients with PCOS.
本研究的目的是阐明微小RNA-27a-3p（miR-27a-3p）在多囊卵巢综合征（PCOS）中的功能。分析了21例接受体外受精周期治疗的PCOS患者和12例正常患者切除的颗粒细胞（GCs），以及17例未经治疗的楔形卵巢切除PCOS样本和13例无PCOS患者的对照卵巢样本中miR-27a-3p的表达。我们发现，与对照组相比，PCOS患者切除的GCs和卵巢中miR-27a-3p的表达均显著增加。对人颗粒细胞瘤细胞系（KGN）进行胰岛素处理导致miR-27a-3p表达下调，这种效应似乎是由信号转导和转录激活因子STAT1和STAT3介导的。miR-27a-3p在KGN细胞中的过表达抑制了SMAD5，进而降低了细胞增殖并促进了细胞凋亡。KGN细胞用胰岛素刺激48小时后，SMAD5蛋白表达增加，细胞凋亡减少。此外，在KGN细胞中敲低/过表达SMAD5可减少/增加细胞数量并促进/抑制细胞凋亡。与正常GCs或KGN细胞相比，胰岛素刺激的PCOS患者分离的原代GCs未表现出miR-27a-3p表达降低。KGN细胞和人PCOS GCs中表达水平的差异可能是由于PCOS中胰岛素抵抗导致GCs中miR-27a-3p表达增加所致。综上所述，我们的数据为miR-27a-3p在PCOS患者GCs功能障碍中的作用提供了证据。
