17β-雌二醇对卵巢颗粒细胞增殖转移的影响及作用机制

Effect of 17β-estradiol on proliferation and metastasis of ovarian granule cells and the mechanism

目的探究17β-雌二醇(17β-E_2)对卵巢颗粒细胞增殖、转移的影响及其作用机制。方法体外培养卵巢颗粒细胞系—KGN细胞,采用MTT法检测17β-E_2对KGN细胞增殖的影响;划痕实验检测17β-E_2单独或和G蛋白偶联雌激素受体1(GPER1)特异性拮抗剂—G15共同处理对KGN细胞转移的影响;分别于17β-E_2单独或和G15共同处理KGN细胞0 min、10 min、30 min及60 min后收集细胞,采用Western Blot法检测细胞ERK1/2的磷酸化水平。结果 MTT结果显示,17β-E_2对KGN细胞增殖无显著影响;划痕实验结果表明,17β-E_2单独处理能够显著抑制KGN细胞的转移,而联合G15处理对细胞转移则无显著影响;Western Blot结果显示,17β-E_2单独处理细胞10 min、30 min及60 min后,细胞ERK1/2磷酸化水平均显著降低;而17β-E_2和G15共同处理细胞不同时间后,细胞ERK1/2磷酸化水平相比于处理前无显著性变化。结论 17β-E_2能够显著抑制KGN细胞的转移,其机制与GPER1介导的雌激素非基因组效应有关。

Objective To explore the effect of 17β-estradiol（17β-E2） on proliferation and migration of ovarian granule cells and the mechanism.Methods Ovarian granule cell line-KGN cells were cultured in vitro.MTT assay was used to detect the effect of 17β-E2 on proliferation of KGN cells;scratch test was used to detect the effects of single application of 17β-E2 and 17β-E2 combined with G15（specific antagonist of G protein-coupled estrogen receptor 1） on migration of KGN cells.After treating KGN cells with single 17β-E2and17β-E2 combined with G15 for 0,10,30,and 60 minutes,the cells were collected.Western Blot was used to detect phosphorylation level of ERK1/2.Results MTT assay showed that 17β-E2 had no effect on proliferation of KGN cells;scratch test showed that single 17β-E2 significantly inhibited migration of KGN cells,but 17β-E2 combined with G15 had no significantly effect on migration of KGN cells;Western Blot showed that after treating KGN cells with single 17β-E2 for 10,30,and 60 minutes,phosphorylation level of ERK1/2 decreased significantly;after treating KGN cells with 17β-E2 combined with G15 for different time periods,there was no statistically significant difference in phosphorylation level of ERK1/2 between before and after treatment.Conclusion 17β-estradiol can inhibit the migration of granule cells significantly,and the mechanism is associated with GPER1 mediated estrogen non-genomic effect.