Zhang Kexin, Wang Xiaomeng, Liu Fang, Lin Hong, Wang Yan, Zhao Min, Wang Xiaofei, Chu Yijing, Xu Lin
Department of Obstetrics, The Affiliated Hospital of Qingdao University, 59 Haier Road, Qingdao, 266000, China.
Qingdao University, Qingdao, China.
J Ovarian Res. 2025 Mar 13;18(1):55. doi: 10.1186/s13048-025-01623-4.
Polycystic ovary syndrome (PCOS) is currently recognized as a condition that affects several systems in the body, including the reproductive, endocrine, and cardiovascular systems. Prevalent among teenagers and women of reproductive age. Prior research has demonstrated an elevation of miR-34a-5p within the follicular fluid (FF) of women of PCOS. Despite this, the precise mechanisms through which miR-34a-5p influences granulosa cells (GC) development and function remain poorly characterized.
Therefore, this study investigates the involvement and pathogenic mechanisms of miR-34a-5p within GCs in the context of PCOS. The human granulosa-like tumor cell line (KGN) got transfected at a control, as well as a miR-34a-5p mimic and inhibitor, respectively. Monitor cellular proliferation in each experimental group. The experimental methods included RT-qPCR, CCK8, flow cytometry and western blotting. Also, the interaction between miR-34a-5p and the particular sequence of JAG1 has been verified using the dual luciferase assay. Further investigation of the connection involving miR-34a-5p and the Notch signaling pathway was conducted using bioinformatics analysis and experimental methods.
The results demonstrated that miR-34a-5p expression was significantly elevated in the serum(p<0. 0001)and FF (p = 0. 0402) of PCOS, whereas its expression in GCs (p = 0. 5522) showed no significant variation. Overexpressing miR-34a-5p caused a decrease in the rate at which KGN cells multiplied and an increase in programmed cell death. Conversely, inhibiting miR-34a-5p resulted in an increase in cell growth and a decrease in programmed cell death. Bioinformatics analysis and experimental results further demonstrated thatmiR-34a-5p interacts with the 3'UTR region of JAG1, leading to a negative regulation of the Jagged1-Notch signaling pathway.
In summary, the miR-34a-5p molecule inhibits the growth of GCs as well as triggers programmed cell death by regulating the Jagged1-Notch signaling pathway. Silencing miR-34a-5p prevents dysfunction in GCs. Our analysis implies that miR-34a-5p is a new molecular site to treat PCOS.
多囊卵巢综合征（PCOS）目前被认为是一种影响身体多个系统的病症，包括生殖、内分泌和心血管系统。在青少年和育龄女性中普遍存在。先前的研究表明，PCOS女性的卵泡液（FF）中miR-34a-5p水平升高。尽管如此，miR-34a-5p影响颗粒细胞（GC）发育和功能的精确机制仍不清楚。
因此，本研究探讨了miR-34a-5p在PCOS背景下在GC中的作用及其致病机制。分别用对照、miR-34a-5p模拟物和抑制剂转染人颗粒细胞瘤细胞系（KGN）。监测每个实验组的细胞增殖情况。实验方法包括RT-qPCR、CCK8、流式细胞术和蛋白质印迹法。此外，使用双荧光素酶报告基因检测验证了miR-34a-5p与JAG1特定序列之间的相互作用。使用生物信息学分析和实验方法进一步研究了miR-34a-5p与Notch信号通路之间的联系。
结果表明，PCOS患者血清（p<0.0001）和FF（p = 0.0402）中miR-34a-5p表达显著升高，而其在GC中的表达（p = 0.5522）无显著变化。过表达miR-34a-5p导致KGN细胞增殖速率降低，程序性细胞死亡增加。相反，抑制miR-34a-5p导致细胞生长增加，程序性细胞死亡减少。生物信息学分析和实验结果进一步表明miR-34a-5p与JAG1的3'UTR区域相互作用，导致Jagged1-Notch信号通路的负调控。
总之，miR-34a-5p分子通过调节Jagged1-Notch信号通路抑制GC生长并引发程序性细胞死亡。沉默miR-34a-5p可预防GC功能障碍。我们的分析表明，miR-34a-5p是治疗PCOS的一个新的分子靶点。
