miR-103a-3p对卵巢颗粒细胞增殖与凋亡的影响

Effects of miR-103a-3p on proliferation and apoptosis in avarian granulosa cells

目的 观察miR-103a-3p对卵巢颗粒细胞增殖与凋亡的影响,初步探讨miR-103a-3p在多囊卵巢综合征(PCOS)中的可能作用。方法 体外培养人卵巢颗粒细胞系KGN细胞株,构建miR-103a-3p高表达的颗粒细胞模型;结合荧光标签及qRT-PCR检测转染效率;MTT法检测细胞活力;EdU染色结合流式细胞术检测细胞增殖;Annexin染色结合流式细胞术、Hoechst法检测细胞凋亡。结果 转染组细胞miR-103a-3p表达明显高于对照组(P<0.05),提示miR-103a-3p已转染至颗粒细胞;与对照组相比,miR-103a-3p过表达组的颗粒细胞活力下降(P<0.05),G0-G1期细胞数量明显增多(P<0.05),而细胞的凋亡率则明显升高(P<0.05)。结论 miR-103a-3p可抑制KGN颗粒细胞增殖,并促进细胞凋亡,可能在PCOS中有着重要作用。

Objective To observe the effect of miR-103a-3p on the proliferation and apoptosis of ovarian granulosa cells, and to preliminarily explore the possible role of miR-103a-3p in polycystic ovary syndrome(PCOS). Methods Human ovarian granulosa cell line KGN was cultured in vitro and transiently transfected to construct a granulosa cell model with high expression of miR-103a-3p;the transfection efficiency was detected by fluorescence labeling and qRT-PCR;cell viability was detected by MTT method;cell proliferation was detected by EdU staining combined with flow cytometry;apoptosis was detected by Annexin staining combined with flow cytometry and Hoechst method. Results The expression of miR-103a-3p in transfected cells was significantly higher than that in control cells(P<0.05);compared with the control group, the viability of granulosa cells in miR-103a-3p overexpression group decreased(P<0.05), the number of cells in G0-G1 phase increased significantly(P<0.05), and the apoptosis rate of cells increased significantly(P<0.05).Conclusion miR-103a-3p can inhibit the proliferation of KGN granulosa cells and promote apoptosis, which may play an important role in PCOS.