Xiao-Ling Ji1, Xia Liu2, Zhe Wang1, Ying-Chun Fang3. 1. Department of Gynecology area 1, Weifang People's Hospital, No. 151, Guangwen Street, Weifang, 261041, Shandong, China. 2. Department of Obstetrics, Affiliated Hospital of Weifang Medical College, Weifang, Shandong, China. 3. Department of Gynecology area 1, Weifang People's Hospital, No. 151, Guangwen Street, Weifang, 261041, Shandong, China. Electronic address: jasmine_fangyc@163.com.
Abstract
OBJECTIVE: 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). METHODS: 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. RESULTS: 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. CONCLUSION: 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.
OBJECTIVE: 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). METHODS: Serum samples were collected from PCOSpatients to detect the expression of ARID1A by qRT-PCR. Then, mouse and humanovarian 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. RESULTS:ARID1A was down-regulated in the serum of PCOSpatients and ovarian GCs from PCOSmice. Human and mouseovarian 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 micePCOS 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 micePCOS. 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. CONCLUSION: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.