Department of Gynecology area 1, Weifang People's Hospital, No. 151, Guangwen Street, Weifang, 261041, Shandong, China.
Department of Obstetrics, Affiliated Hospital of Weifang Medical College, Weifang, Shandong, China.
Ann Endocrinol (Paris). 2020 Dec;81(6):521-529. doi: 10.1016/j.ando.2020.11.008. Epub 2020 Dec 5.
The purpose of the present study was to clarify the expression of ARID1A in polycystic ovary syndrome (PCOS) and its effect on ovarian granulosa cells (GCs).
Serum samples were collected from PCOS patients to detect the expression of ARID1A by qRT-PCR. Then, mouse and human ovarian GCs were isolated and divided into several groups according to difference in transfection, and the following experiments were performed: MTT assay, flow cytometry, qRT-PCR, radioimmunoassay, and Western blotting.
ARID1A was down-regulated in the serum of PCOS patients and ovarian GCs from PCOS mice. Human and mouse ovarian GCs in the ARID1A group and in cells that were exposed to LY294002, a PI3/Akt pathway inhibitor, showed decreased proliferation and increased apoptosis compared to those in the mock group, and a higher percentage of G0/G1 phase with a lower percentage of S phase or G2/M. Moreover, the expression of steroid metabolism-related genes (3βHSD,Cyp11a1, StAR and Cyp19a1) in both human and mice PCOS GCs was down-regulatedresulting in lower estradiol (E2) and progesterone (P) 48h accumulation. In addition, protein expression of cleaved caspase-3, a main executor of apoptosis, was increased while expression of p-Akt/Akt and cyclin D1 was decreased in GCs from human and mice PCOS. However, the levels of the above indicators in the si-ARID1A group showed inverse changes. Furthermore, LY29400 treatment could reverse the effect of si-ARID1A on the ovarian GCs.
ARID1A was down-regulated in GCs cells form PCOS women and from PCOS animal models, while ARID1A overexpression can suppress the PI3K/Akt pathway to inhibit proliferation and promote apoptosis in ovarian granulosa cells.
本研究旨在阐明 ARID1A 在多囊卵巢综合征（PCOS）中的表达及其对卵巢颗粒细胞（GCs）的影响。
收集 PCOS 患者的血清样本，通过 qRT-PCR 检测 ARID1A 的表达。然后，分离小鼠和人卵巢 GCs，并根据转染的差异将其分为几组，进行以下实验：MTT 测定、流式细胞术、qRT-PCR、放射免疫测定和 Western blot。
PCOS 患者血清和 PCOS 小鼠卵巢 GCs 中 ARID1A 表达下调。与模拟组相比，ARID1A 组和暴露于 PI3/Akt 通路抑制剂 LY294002 的人源和鼠源卵巢 GCs 的增殖减少，凋亡增加，G0/G1 期比例升高，S 期或 G2/M 期比例降低。此外，人源和鼠源 PCOS GCs 中类固醇代谢相关基因（3βHSD、Cyp11a1、StAR 和 Cyp19a1）的表达下调，导致雌二醇（E2）和孕酮（P）48 小时积累减少。此外，人源和鼠源 PCOS GCs 中 cleaved caspase-3 的蛋白表达增加，凋亡的主要执行者，而 p-Akt/Akt 和 cyclin D1 的表达减少。然而，si-ARID1A 组的上述指标水平呈相反变化。此外，LY294002 处理可逆转 si-ARID1A 对卵巢 GCs 的作用。
在来自 PCOS 女性和 PCOS 动物模型的 GCs 细胞中，ARID1A 表达下调，而 ARID1A 过表达可抑制 PI3K/Akt 通路，抑制卵巢颗粒细胞增殖，促进凋亡。
