Reproductive Medical Centre, Renmin Hospital of Wuhan University, Wuhan, China.
J Cell Mol Med. 2022 Mar;26(6):1817-1825. doi: 10.1111/jcmm.16231. Epub 2020 Dec 28.
Polycystic Ovary Syndrome (PCOS) is a kind of endocrine disorder which is prevalent in adult women, so exploring more biomarkers for PCOS is imperative. Recently, circular RNA and microRNA are confirmed to be related with PCOS development. Whether circular RNA ASPH (circASPH) is involved in PCOS need to be studied further. We utilized RT-qPCR to measure the expression levels of circASPH, miR-375 and MAP2K6 in PCOS patients and normal group. The effects of circASPH and miR-375 on KGN cells proliferation and apoptosis were observed by CCK-8 assay, EdU incorporation assay and apoptosis assay, separately. Then Dual-luciferase reporter assay was carried out to verify the circASPH/miR375 axis and miR375/MAP2K6 axis. The interaction between circASPH and MAP2K6 were detected with the support of RT-qPCR and Western blot. We found circASPH and MAP2K6 were both over-expressed in PCOS patients, while miR-375 was in the opposite direction. Moreover, miR-375 was negatively regulated by circASPH, while MAP2K6 was positively regulated by circASPH. In addition, circASPH directly targeted miR-375, which targeted MAP2K6. More than that, the knockdown of circASPH repressed KGN cells proliferation and enhanced apoptosis, while the silence of miR-375 reversed the above effects. In conclusion, circASPH promotes KGN cells proliferation through miR-375/MAP2K6 axis in PCOS, and they are thought-provoking biomarkers for PCOS diagnosis and therapy.
多囊卵巢综合征(PCOS)是一种常见于成年女性的内分泌疾病,因此探索更多的 PCOS 生物标志物是当务之急。最近,环状 RNA 和 microRNA 被证实与 PCOS 的发展有关。环状 RNA ASPH(circASPH)是否参与 PCOS 还需要进一步研究。我们利用 RT-qPCR 测量了 PCOS 患者和正常组中 circASPH、miR-375 和 MAP2K6 的表达水平。通过 CCK-8 测定、EdU 掺入测定和凋亡测定分别观察 circASPH 和 miR-375 对 KGN 细胞增殖和凋亡的影响。然后进行双荧光素酶报告基因检测验证 circASPH/miR375 轴和 miR375/MAP2K6 轴。利用 RT-qPCR 和 Western blot 检测 circASPH 和 MAP2K6 之间的相互作用。我们发现 circASPH 和 MAP2K6 在 PCOS 患者中均过度表达,而 miR-375 则相反。此外,miR-375 被 circASPH 负调控,而 MAP2K6 被 circASPH 正调控。另外,circASPH 直接靶向 miR-375,而 miR-375 靶向 MAP2K6。不仅如此,circASPH 的敲低抑制了 KGN 细胞的增殖并增强了凋亡,而 miR-375 的沉默逆转了上述作用。综上所述,circASPH 通过 miR-375/MAP2K6 轴促进 PCOS 中 KGN 细胞的增殖,它们是 PCOS 诊断和治疗有意义的生物标志物。