摘要
目的探讨微小RNA-1270(miR-1270)及其靶基因细胞色素P45019A1(CYP19A1)在多囊卵巢综合征(PCOS)患者卵丘颗粒细胞中的表达,并评价其临床意义。方法回顾性分析2019年4月至2020年12月期间行IVF或ICSI助孕的PCOS患者(PCOS组)和无PCOS的不孕患者(对照组)的临床资料,记录两组患者的基本临床资料和促排卵情况。并收集两组患者的卵丘颗粒细胞,采用实时荧光定量PCR(qRT-PCR)检测卵丘颗粒细胞中miR-1270与CYP19A1的表达。双荧光素酶报告基因系统检测miR-1270是否靶向结合CYP19A1。将miR-1270的模拟物或抑制剂转染至人卵巢颗粒细胞瘤细胞KGN,采用qRT-PCR和蛋白免疫印迹检测CYP19A1的表达变化,采用ELISA测定KGN细胞培养液上清中雌二醇(E 2)的浓度。结果PCOS组不孕患者的窦卵泡数、体重指数(BMI)、抗苗勒管激素(AMH)、LH、LH/FSH、催乳素(PRL)、睾酮(T)均显著高于对照组(P<0.05),获卵数亦显著高于对照组(P<0.05)。与对照组相比,PCOS组卵丘颗粒细胞中miR-1270表达量显著降低(P<0.05),CYP19A1表达量显著增加(P<0.05)。双荧光素酶报告基因实验证实了CYP19A1和miR-1270之间是直接结合的,并且miR-1270通过靶向结合CYP19A1抑制E 2合成。结论miR-1270靶向负调控CYP19A1的表达,影响PCOS患者卵丘颗粒细胞中E 2的生成,为PCOS患者卵泡发育障碍提出了可能的分子机制。
Objective:To investigate the expression of miR-1270 and its target gene aromatase cytochrome P450(CYP19A1)in granulosa cells of patients with polycystic ovary syndrome(PCOS),and to explore its clinical significance.Methods:The clinical data of PCOS patients(PCOS group)and infertile patients without PCOS(control group)who underwent IVF or ICSI from April 2019 to December 2020 were retrospectively analyzed.All patients’basic clinical data and ovulation induction were recorded.The granulosa cells were collected from the patients of the two groups.The qRT-PCR was used to identity the expression of miR-1270 and CYP19A1 of cumulus granulosa cells.Dual-luciferase reporter gene system was used to detect whether miR-1270 targeted CYP19A1.The mimic or inhibitor of miR-1270 was transfected in KGN cells(human ovary granulosa tumor cells).CYP19A1 mRNA and protein expression were measured by qRT-PCR and Western blot.The concentration of E 2 in KGN cells culture medium was measured by ELISA.Results:The antral follicles counts,body mass index(BMI),anti-Mullerian hormone(AMH),LH,LH/FSH,prolactin(PRL)and testosterone(T)in PCOS group were significantly higher than those in control group(P<0.05),and the number of oocytes retrieved was also significantly higher than that of the control group(P<0.05).The expression of miR-1270 was significantly decreased(P<0.05)in granulosa cells of patients with PCOS,while CYP19A1 was significantly increased(P<0.05).The direct combination between miR-1270 and CYP19A1 was confirmed by dual-luciferase reporter assays.Moreover,miR-1270 inhibited E 2 synthesis by targeting CYP19A1.Conclusions:MiR-1270 negatively regulated the expression of CYP19A1 and affected estradiol synthesis in cumulus granulosa cells of patients with PCOS.This study offered a possible mechanism of follicle development arrest in patients with PCOS.
作者
张慧敏
朱文倩
马静
徐仰英
张玉
沈伟
郝翠芳
ZHANG Hui-min;ZHU Wen-qian;MA Jing;XU Yang-ying;ZHANG Yu;SHEN Wei;HAO Cui-fang(Medical College of Qingdao University,Qingdao 266071;Department of Clinical Medicine,Binzhou Medical University,Yantai 264010;Centre for Reproductive Medicine,Qingdao Woman and Children’s Hospital,Qingdao University,Qingdao 266011;College of Life Science and Technology,Qingdao Agricultural University,Qingdao266109;Centre for Reproductive Medicine,Yantai Yuhuangding Hospital Affiliated to Qingdao University,Yantai 264000)
出处
《生殖医学杂志》
CAS
2021年第5期649-657,共9页
Journal of Reproductive Medicine
基金
国家自然科学基金面上项目(81671416)
青岛市医疗卫生重点学科建设项目。
