天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (7): 673-677.doi: 10.11958/20210042

• 细胞与分子生物学 •    下一篇

MicroRNA-141通过PI3K/Akt/mTOR信号通路促进PCOS卵巢颗粒细胞增殖的机制研究

陈娇，张妍，钟媛媛   

  1. 华中科技大学同济医学院附属武汉儿童医院（武汉市妇幼保健院）妇产科（邮编430015）
  • 收稿日期:2021-01-11 修回日期:2021-01-29 出版日期:2021-07-15 发布日期:2021-07-12
  • 作者简介:陈娇（1980），女，学士，住院医师，主要从事多囊卵巢综合征基础与临床研究。E-mail：cjfck3333@126.com
  • 基金资助:
    武汉市卫健委科研项目（WX15B10）

MicroRNA-141 promotes the proliferation of ovarian granulosa cells in polycystic ovary syndrome through PI3K/Akt/mTOR signaling pathway

CHEN Jiao, ZHANG Yan, ZHONG Yuan-yuan   

  1. Department of Obstetrics and Gynecology, Wuhan Children's Hospital (Wuhan Maternal and Child Healthcare Hospital), Tongji Medical College, Huazhong University of Science & Technology, Wuhan 430015, China
  • Received:2021-01-11 Revised:2021-01-29 Published:2021-07-15 Online:2021-07-12

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摘要： 目的　研究microRNA-141（miR-141）对卵巢颗粒细胞增殖的影响，并探讨其在多囊卵巢综合征（PCOS）发生发展中的作用机制。方法　采用实时荧光定量PCR检测20例PCOS患者的卵巢组织、20例对照组的正常卵巢组织、人卵巢颗粒细胞KGN及人正常卵巢上皮细胞IOSE80中miR-141 mRNA表达水平；KGN细胞分为miR-141 minics组、LY294002+miR-141 minics组、雷帕霉素（rapamycin）+miR-141 minics组、NC组、对照组。采用MTT法和平板克隆实验检测各组细胞增殖和克隆形成能力，采用Western blot法检测各组细胞磷脂酰肌醇3-激酶（PI3K）/蛋白激酶（Akt）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路相关蛋白表达水平。结果　PCOS卵巢组织miR-141 mRNA表达水平显著高于正常卵巢组织，人卵巢颗粒细胞KGN miR-141 mRNA表达水平高于人正常卵巢上皮细胞IOSE80（P＜0.05）；miR-141 minics组、LY294002+miR-141 minics组及rapamycin+miR-141 minics组miR-141 mRNA表达水平均高于NC组和对照组（P＜0.05）；LY294002+miR-141 minics组及rapamycin+miR-141 minics组转染后24、48、72、96 h OD值和克隆形成率均低于miR-141 minics组，但高于NC组和对照组（P＜0.05）；LY294002+miR-141 minics组转染后p-Akt、p-mTOR蛋白表达水平低于miR-141 minics组，但高于NC组、对照组（P＜0.05）。rapamycin+miR-141 minics组转染后p-mTOR蛋白表达水平低于miR-141 minics组，高于NC组、对照组（P＜0.05）。结论　miR-141可通过激活PI3K/Akt/mTOR信号通路促进卵巢颗粒细胞的增殖。

关键词: 多囊卵巢综合征, 磷酸肌醇3-激酶类, 粒层细胞, 蛋白激酶B, miR-141, 卵巢颗粒细胞, PI3K/Akt/mTOR信号通路

Abstract: Objective　To study the effect of microRNA-141 (miR-141) on the proliferation of ovarian granulosa cells and to explore its mechanism in the occurrence and development of polycystic ovary syndrome (PCOS). Methods　qPCR was used to detect the expression levels of miR-141 mRNA in 20 samples PCOS ovarian tissues, 20 normal ovarian tissue specimens, human ovarian granulosa cells KGN and human normal ovarian epithelial cells IOSSE80. KGN cells were divided into miR-141 minics group, LY294002+miR-141 minics group, rapamycin+miR-141 minics group, NC group and control group. The MTT method and the plate cloning experiment method were used to detect the cell proliferation and clone formation ability of each group, and the Western blot assay was used to detect the phosphatidylinositol 3-kinase (PI3K)/protein kinase (Akt)/mammalian rapamycin target protein (mTOR) signaling pathway related protein expression levels. Results　The expression level of miR-141 mRNA was significantly higher in PCOS ovarian tissue than that in normal ovarian tissue, and the expression level of miR-141 mRNA was significantly higher in human ovarian granulosa cells KGN than that in normal ovarian epithelial cells IOSE80 (P＜0.05). miR-141 mRNA expression levels were significantly higher in miR-141 minics group, LY294002+miR-141 minics group and rapamycin+miR-141 minics group than those in NC group and control group (P＜0.05). After transfection for 24, 48, 72 and 96 h, the OD values and clone formation rates were lower in LY294002+miR-141 minics group and rapamycin+miR-141 minics group than those of miR-141 minics group, but higher than those of NC group and control group (P＜0.05). After transfection, the expression levels of p-Akt and p-mTOR were lower in LY294002+miR-141 minics group than those in miR-141 minics group, but higher than those in NC group and control group (P＜0.05). The expression level of p-mTOR protein was significantly lower in rapamycin+miR-141 minics group than that in miR-141 minics group, and higher than that in NC group and control group (P＜0.05). Conclusion　miR-141 can promote proliferation of ovarian granulosa cells by activating the PI3K/Akt/mTOR signaling pathway.

Key words: polycystic ovary syndrome, phosphatidylinositol 3-kinases, granulosa cells, protein kinase B, miR-141, ovarian granulosa cells, PI3K/Akt/mTOR signaling pathway