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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 investigate the effect of 17β-estradiol (17β-E_2) on the proliferation and metastasis of ovarian granulosa cells and its mechanism. Methods The ovarian granulosa cell line-KGN was cultured in vitro. The effect of 17β-E 2 on the proliferation of KGN cells was detected by MTT assay. Scratch assay was used to detect the specific antagonism of 17β-E 2 and G protein-coupled estrogen receptor 1 (GPER1) KG15 cells were treated with 17β-E 2 alone or with G15 for 0 min, 10 min, 30 min and 60 min respectively. Western blotting was used to detect the expression of ERK1 / 2 Phosphorylation levels. Results MTT results showed that 17β-E 2 had no significant effect on the proliferation of KGN cells. Scratch test results showed that 17β-E 2 alone could significantly inhibit the metastasis of KGN cells, while the combination of G15 had no significant effect on the cell metastasis. Western Blot results The results showed that the phosphorylation level of ERK1 / 2 in cells treated with 17β-E 2 alone for 10 min, 30 min and 60 min decreased significantly. However, when the cells were treated with 17β-E 2 and G 15 for different time, ERK1 / 2 phosphorylation No significant change than before treatment. Conclusion 17β-E 2 can significantly inhibit KGN cell metastasis, and its mechanism is related to GPER1-mediated estrogen non-genomic effects.