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Circ_0005925通过靶向miR-324-3p上调多囊卵巢综合征中MAP2K6促进颗粒细胞生长。

Circ_0005925 Promotes Granulosa Cell Growth by Targeting MiR-324-3p to Upregulate MAP2K6 in Polycystic Ovary Syndrome.

作者信息

Tu Peng, Yan Sujuan, Zhang Fengping

机构信息

Department of Reproductive Medicine, The First People's Hospital of Jingmen, No.168 Xiangshan Avenue, Duodao District, Jingmen, 448000, Hubei, China.

出版信息

Biochem Genet. 2023 Feb;61(1):21-34. doi: 10.1007/s10528-022-10238-z. Epub 2022 Jun 11.

Abstract

Circular RNA has been reported to be involved in the development of polycystic ovary syndrome (PCOS), but the role of circ_0005925 in the progression of PCOS is unclear. The expression levels of circ_0005925, microRNA (miR)-324-3p, and MAPK kinase 6 (MAP2K6) were examined by quantitative real-time PCR. Cell proliferation and apoptosis were determined using cell counting kit 8 assay, EdU assay, colony formation assay, and flow cytometry. Western blot analysis was used to detect the protein expression of apoptosis markers and MAP2K6. RNA interaction was verified by dual-luciferase reporter assay and RNA pull-down assay. Circ_0005925 was upregulated in GCs from PCOS patients, as well as in KGN and SVOG cells. Circ_0005925 knockdown repressed GCs proliferation and promoted apoptosis. MiR-324-3p was downregulated in PCOS patients, and it was sponged by circ_0005925. The regulation of circ_0005925 knockdown on GCs growth was abolished by miR-324-3p inhibitor. MAP2K6 was targeted by miR-324-3p, and its expression was positively regulated by circ_0005925. MiR-324-3p inhibited GCs proliferation and increased apoptosis by targeting MAP2K6. Collectively, our findings suggested that circ_0005925 promoted GCs growth through miR-324-3p/MAP2K6, which provided a promising therapeutic target for PCOS.

摘要

据报道,环状RNA参与了多囊卵巢综合征(PCOS)的发生发展,但circ_0005925在PCOS进展中的作用尚不清楚。通过定量实时PCR检测circ_0005925、微小RNA(miR)-324-3p和丝裂原活化蛋白激酶激酶6(MAP2K6)的表达水平。使用细胞计数试剂盒8检测、EdU检测、集落形成检测和流式细胞术来测定细胞增殖和凋亡。蛋白质印迹分析用于检测凋亡标志物和MAP2K6的蛋白表达。通过双荧光素酶报告基因检测和RNA下拉检测验证RNA相互作用。circ_0005925在PCOS患者的颗粒细胞(GCs)以及KGN和SVOG细胞中上调。circ_0005925敲低可抑制GCs增殖并促进凋亡。miR-324-3p在PCOS患者中下调,并且被circ_0005925海绵化。miR-324-3p抑制剂消除了circ_0005925敲低对GCs生长的调节作用。MAP2K6是miR-324-3p的靶标,其表达受到circ_0005925的正向调节。miR-324-3p通过靶向MAP2K6抑制GCs增殖并增加凋亡。总的来说,我们的研究结果表明,circ_0005925通过miR-324-3p/MAP2K6促进GCs生长,这为PCOS提供了一个有前景的治疗靶点。

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