Reproductive Medicine Center, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
Department of Ophthalmology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
J Cell Mol Med. 2020 Oct;24(20):11874-11882. doi: 10.1111/jcmm.15807. Epub 2020 Sep 1.
The pathophysiology of polycystic ovary syndrome (PCOS) is characterized by granulosa cell (GC) dysfunction. m A modification affects GC function in patients with premature ovarian insufficiency (POI), but the role of m A modification in PCOS is unknown. The purpose of the prospective comparative study was to analyse the m A profile of the luteinized GCs from normovulatory women and non-obese PCOS patients following controlled ovarian hyperstimulation. RNA m A methylation levels were measured by m A quantification assay in the luteinized GCs of the controls and PCOS patients. Then, m A profiles were analysed by methylated RNA immunoprecipitation sequencing (MeRIP-seq). We reported that the m A level was increased in the luteinized GCs of PCOS patients. Comparative analysis revealed differences between the m A profiles from the luteinized GC of the controls and PCOS patients. We identified FOXO3 mRNA with reduced m A modification in the luteinized GCs of PCOS patients. Selectively knocking down m A methyltransferases or demethylases altered expression of FOXO3 in the luteinized GCs from the controls, but did not in PCOS patients. These suggested an absence of m A-mediated transcription of FOXO3 in the luteinized GCs of PCOS patients. Furthermore, we demonstrated that the involvement of m A in the stability of the FOXO3 mRNA that is regulated via a putative methylation site in the 3'-UTR only in the luteinized GCs of the controls. In summary, our findings showed that altered m A modification was involved in up-regulated expression of FOXO3 mRNA in the luteinized GCs from non-obese PCOS patients following controlled ovarian hyperstimulation.
多囊卵巢综合征（PCOS）的病理生理学特征是颗粒细胞（GC）功能障碍。m A 修饰会影响卵巢早衰（POI）患者的 GC 功能，但 m A 修饰在 PCOS 中的作用尚不清楚。本前瞻性对照研究的目的是分析在控制性卵巢过度刺激后，正常排卵的妇女和非肥胖 PCOS 患者的黄体化 GC 中的 m A 谱。通过 m A 定量测定法测量对照和 PCOS 患者黄体化 GC 中的 RNA m A 甲基化水平。然后，通过甲基化 RNA 免疫沉淀测序（MeRIP-seq）分析 m A 谱。我们报道 PCOS 患者黄体化 GC 中的 m A 水平增加。比较分析显示了对照和 PCOS 患者黄体化 GC 的 m A 谱之间的差异。我们发现 FOXO3 mRNA 的 m A 修饰减少，这是 PCOS 患者黄体化 GC 中的特征。选择性敲低 m A 甲基转移酶或去甲基酶改变了对照黄体化 GC 中 FOXO3 的表达，但在 PCOS 患者中没有。这表明在 PCOS 患者的黄体化 GC 中，FOXO3 的转录不受 m A 介导。此外，我们证明 m A 参与了在黄体化 GC 中受 3'-UTR 中假定甲基化位点调节的 FOXO3 mRNA 的稳定性，这仅在对照的黄体化 GC 中起作用。总之，我们的研究结果表明，在控制性卵巢过度刺激后，非肥胖 PCOS 患者的黄体化 GC 中，改变的 m A 修饰参与了 FOXO3 mRNA 的上调表达。
