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目的探讨G蛋白耦联受体30(GPR30)介导的雌激素非基因效应促乳腺癌细胞侵袭的作用及其机制。方法体外培养乳腺癌细胞株SK-BR-3(GPR30+,ERα-/ERβ-),分别给予不同浓度的17β-雌二醇(E2)和GPR30特异性激动剂(G1)处理不同时间,应用免疫印迹法观察黏着斑激酶(FAK)的磷酸化;转染GPR30 si RNA抑制GPR30表达后,采用Transwell侵袭小室法观察E2和G1对细胞迁移和侵袭的影响。结果与对照组比较,不同浓度的E2(10-10、10-9、10-8、10-7、10-6mol/L)或G1(10-8、10-7、10-6 mol/L)处理SK-BR-3细胞20 min,均能促进FAK的磷酸化。转染GPR30 si RNA 48h抑制GPR30表达后,E2、G1促进FAK磷酸化的效果均明显减弱。E2、G1均可明显促进SK-BR-3细胞的侵袭。而转染si RNA GPR30抑制GPR30表达后,E2、和G1促进细胞侵袭的能力明显减弱。结论雌激素可通过作用于GPR30激活非基因效应诱导FAK的磷酸化,增加乳腺癌细胞的侵袭能力。
Objective To investigate the role of G-protein coupled receptor 30 (GPR30) -mediated estrogen non-gene effect in breast cancer cell invasion and its mechanism. Methods The breast cancer cell lines SK-BR-3 (GPR30 +, ERα- / ERβ-) were cultured in vitro and treated with different concentrations of 17β-estradiol (E2) and GPR30-specific agonists The phosphorylation of focal adhesion kinase (FAK) was detected by Western blotting. GPR30 si RNA was transfected into GPR30 siRNA to inhibit GPR30 expression. Transwell invasion chamber was used to observe the effect of E2 and G1 on cell migration and invasion. Results Compared with the control group, E2 (10-10,10-9,10-8,10-7,10-6 mol / L) or G1 (10-8,10-7,10-6 mol / L) ) Treatment of SK-BR-3 cells for 20 min, can promote FAK phosphorylation. GPR30 si RNA transfected 48h inhibition of GPR30 expression, E2, G1 FAK phosphorylation effect was significantly reduced. E2, G1 can significantly promote the invasion of SK-BR-3 cells. The transfected with si RNAi GPR30 inhibited the expression of GPR30, E2, and G1 to promote cell invasion ability was significantly weakened. Conclusion Estrogen can induce the phosphorylation of FAK by activating non-gene effect on GPR30 and increase the invasiveness of breast cancer cells.