4-羟基他莫昔芬通过雌激素受体GPR30激活ezrin蛋白促进人乳腺癌MCF-7细胞迁移

4-Hydroxytamoxifen-stimulated activation of ezrin is mediated via G-protein-coupled receptor 30 and promotes human breast cancer MCF-7 cell migration

目的:观察4-羟基他莫昔芬(OHT)对人乳腺癌细胞株MCF-7迁移的影响并探讨其机制。方法:贴壁培养雌激素受体阳性人乳腺癌细胞株MCF-7,细胞划痕愈合实验观察OHT对MCF-7细胞迁移的影响,Western blotting检测c-Src/p-Src和ezrin/p-ezrin蛋白的表达。结果:(1)与对照组相比,单独给予MCF-7细胞雌二醇(E2)和OHT都能促进细胞迁移,OHT不能抑制E2对MCF-7细胞的促迁移效应。(2)OHT促进MCF-7细胞迁移最大效应浓度约为5μmol/L,在6 h即能观察到明显促迁移效用。(3)OHT和雌激素受体G蛋白偶联受体30(GPR30)激动剂G1都能明显增加p-Src和p-ezrin蛋白表达。(4)分别用G15和PP2阻断GPR30和Src后,OHT激活ezrin作用都能被阻断。结论:OHT可能通过结合GPR30后激活Src蛋白,进而磷酸化激活ezrin,介导细胞骨架重构并促进MCF-7细胞迁移。

AIM： To investigate the effect of 4-hydroxytamoxifen （OHT） on the migration of human breast cancer cell line MCF-7. METHODS： The cell migration ability was assayed by scratch healing experiment. The protein expression of Src, p-Src, ezrin and p-ezrin were examined by Western blotting. RESULTS ： The results of scratch healing experiment confirmed that both OHT and estradiol （E2 ） promoted MCF-7 cell migration and the E2-enhanced the cells migration was not inhibited by OHT. The most effective concentration of OHT that enhanced cell migration was 5 μ mol/L. Significant promotion of the cell migration was observed at 6 h after OHT treatment. Increased p-ezrin and p-Src expression was observed after treatment with OHT and G-protein-coupled receptor 30 （GPR30） agonists G1. The expression of p-ezrin after OHT treatment was inhibited by G15 （GPR30 blocker） or PP2 （Src inhibitor）. CONCLUSION： 4-Hydroxytamoxifen promotes MCF-7 cell migration by activation of ezrin via GPR30 and c-Sre.