Tu Peng, Yan Sujuan, Zhang Fengping
Department of Reproductive Medicine, The First People's Hospital of Jingmen, No.168 Xiangshan Avenue, Duodao District, Jingmen, 448000, Hubei, China.
Biochem Genet. 2023 Feb;61(1):21-34. doi: 10.1007/s10528-022-10238-z. Epub 2022 Jun 11.
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.
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。
