Reproductive Medicine Center, Tongji Hospital, Tongji Medicine College, Huazhong University of Science and Technology, Jiefang Avenue 1095#, Wuhan, 430030, People's Republic of China.
Reprod Sci. 2024 Nov;31(11):3498-3511. doi: 10.1007/s43032-024-01632-6. Epub 2024 Jul 12.
In this study, we aimed to investigate the molecular mechanisms of RNA N6-methyladenosine (m6A) modification and how its associated proteins affect granulosa cell aging. A granulosa cell senescence model was constructed to detect the differences in total RNA m6A modification levels and the expression of related enzymes. Changes in downstream molecular expression and the effects on the cellular senescence phenotype were explored by repeatedly knocking down and overexpressing the key genes fat mass and obesity-associated protein (FTO), YT521-B homology domain family member 2 (YTHDF2), and matrix metalloproteinase-2 (MMP2). There was an increased total RNA m6A modification and decreased expression of the demethylase FTO and target gene MMP2 in senescent granulosa cells. FTO and MMP2 knockdown promoted granulosa cell senescence, whereas FTO and MMP2 overexpression retarded it. YTHDF2 and FTO can bind to the messenger RNA of MMP2. The extracellular signal-regulated kinase (ERK) pathway, which is downstream of MMP2, retarded the process of granulosa cell senescence through ERK activators. In granulosa cells, FTO can regulate the expression of MMP2 in an m6A-YTHDF2-dependent manner, influencing the activation status of the ERK pathway and contributing to the aging process of granulosa cells.
在这项研究中，我们旨在探讨 RNA N6-甲基腺苷（m6A）修饰的分子机制，以及其相关蛋白如何影响颗粒细胞衰老。构建了颗粒细胞衰老模型，以检测总 RNA m6A 修饰水平和相关酶表达的差异。通过反复敲低和过表达关键基因脂肪量和肥胖相关蛋白（FTO）、Y 碱基 T 碱基 521 同源结构域家族成员 2（YTHDF2）和基质金属蛋白酶-2（MMP2），探讨下游分子表达的变化及其对细胞衰老表型的影响。衰老的颗粒细胞中总 RNA m6A 修饰增加，去甲基酶 FTO 和靶基因 MMP2 的表达降低。FTO 和 MMP2 的敲低促进了颗粒细胞衰老，而过表达则延缓了这一过程。YTHDF2 和 FTO 可以与 MMP2 的信使 RNA 结合。MMP2 的下游细胞外信号调节激酶（ERK）通路通过 ERK 激活剂延缓了颗粒细胞衰老过程。在颗粒细胞中，FTO 可以以 m6A-YTHDF2 依赖的方式调节 MMP2 的表达，影响 ERK 通路的激活状态，并促进颗粒细胞的衰老过程。
