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.

中文翻译：

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ARID1A在多囊卵巢综合征中的表达及其对卵巢颗粒细胞增殖和凋亡的影响

客观的

本研究的目的是阐明 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。

方法

从 PCOS 患者收集血清样本，通过 qRT-PCR 检测 ARID1A 的表达。然后，根据转染的差异将小鼠和人卵巢GCs分离并分成几组，并进行以下实验：MTT测定、流式细胞术、qRT-PCR、放射免疫测定和Western印迹。

结果

ARID1A在 PCOS 患者的血清和 PCOS 小鼠的卵巢 GC 中下调。与模拟组相比，ARID1A 组和暴露于 LY294002（一种 PI3/Akt 通路抑制剂）的细胞中的人和小鼠卵巢 GC 显示出增殖减少和凋亡增加，并且 G0/G1 期百分比较高， S 期或 G2/M 的百分比较低。此外，类固醇代谢相关基因（3βHSD、 Cyp11a1、StAR和Cyp19a1）的表达) 在人和小鼠中，PCOS GCs 被下调，导致雌二醇 (E2) 和孕酮 (P) 48 小时积累降低。此外，在人类和小鼠 PCOS 的 GC 中，裂解的 caspase-3（细胞凋亡的主要执行者）的蛋白质表达增加，而 p-Akt/Akt 和细胞周期蛋白 D1 的表达降低。而si-ARID1A组上述指标的水平则呈现出相反的变化。此外，LY29400 治疗可以逆转 si-ARID1A 对卵巢 GC 的影响。

结论

ARID1A 在来自 PCOS 女性和 PCOS 动物模型的 GCs 细胞中下调，而 ARID1A 过表达可以抑制 PI3K/Akt 通路以抑制卵巢颗粒细胞的增殖和促进凋亡。