1 汉川市人民医院内分泌科，湖北 汉川 431600
2 汉川市人民医院生殖科，湖北 汉川 431600
3 汉川市人民医院妇产科，湖北 汉川 431600
目的：研究microRNA-184(miR-184)在多囊卵巢综合征(polycystic ovary syndrome，PCOS)中的作 用，并初步探讨其潜在的作用机制。方法：采用qRT-PCR检测24例PCOS卵巢组织标本、20例正 常卵巢组织标本及人卵巢颗粒细胞KGN和人正常卵巢上皮细胞IOSE80中miR-184的表达情况。通 过细胞转染法抑制KGN细胞中miR-184的表达，MTT法检测细胞增殖能力，平板克隆实验检测细 胞克隆形成能力，Western印迹法检测细胞中p-p38 MAPK和p-ERK1/2蛋白表达水平。以终浓度为 20 μmol/L的MAPK信号通路特异性抑制剂SB203580处理下调miR-184的KGN细胞，按照上述方法 检测对细胞增殖的影响。结果：PCOS卵巢组织和KGN细胞中miR-184的表达显著上调。在KGN 细胞中转染miR-184抑制剂可显著抑制miR-184的表达。下调miR-184的表达后，KGN细胞增殖 和细胞克隆形成能力显著降低，细胞中p-p38 MAPK和p-ERK1/2蛋白表达水平显著下降，与对照 组相比，差异有统计学意义(P<0.05)。抑制剂SB203580处理下调miR-184的KGN细胞，细胞增殖 和细胞克隆形成能力显著降低，与单纯下调miR-184的细胞相比，差异有统计学意义(P<0.05)。 结论：MiR-184能够促进卵巢颗粒细胞的增殖，其作用机制与MAPK信号通路的激活有关，可为 PCOS的治疗提供潜在的作用靶点。
MiR-184 promotes the proliferation of ovarian granulosa cells in polycystic ovary syndrome by MAPK signaling pathway
This work supported by the Project of Hubei Provincial Department of Science and Technology, China (EK2017D200002000012).
Objective: To investigate the role of microRNA-184 (miR-184) in polycystic ovary syndrome (PCOS) and to explore its potential mechanism. Methods: Fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the miR-184 expression in 24 cases of PCOS ovarian tissue specimens, 20 normal ovarian tissue specimens, human ovarian granulosa cells KGN and human normal ovarian epithelial cells IOSE80. The expression of miR-184 in KGN cells was inhibited by cell transfection. The proliferation of cells was detected by MTT assay. The ability of cell clone formation was detected by plate cloning assay. The p-p38 MAPK and p-ERK1/2 were detected by Western blot. The KGN cells of down-regulated miR-184 were treated with MAPK signaling pathway inhibitor SB203580 with a final concentration of 20 μmol/L, and the effect on cell proliferation was detected by the above method. Results: The expression of miR-184 was significantly up-regulated in ovarian tissue and KGN cells of polycystic ovary syndrome. Transfection of miR-184 inhibitor in KGN cells significantly inhibited the expression of miR-184. After down-regulating the expression of miR-184, the proliferation of KGN cells and the ability of cell clone formation were significantly decreased, and the expression levels of p-p38 MAPK and p-ERK1/2 protein in the cells were significantly decreased. Compared with the control group, the difference was statistically significant (P<0.05). Inhibitor SB203580 down-regulated miR-184 expression in KGN cells, and the cell proliferation and cell clone formation ability were significantly decreased, compared with the cells directly down-regulated miR-184, the difference was statistically significant (P<0.05). Conclusion: MiR-184 can promote the proliferation of ovarian granulosa cells, and its mechanism is related to the activation of MAPK signaling pathway, which provides a potential target for the treatment of PCOS.
miR-184; MAPK signaling pathway; polycystic ovary syndrome; ovarian granulosa cells