GPER介导的丹参酮ⅡA抗三阴性乳腺癌细胞MDA-MB-231迁移作用机制研究

Investigation of Anti-migration Effect of Tanshinone ⅡA Mediated by GPER on Triple-negative Breast Cancer Cells MDA-MB-231

目的:基于体外细胞实验,探索丹参酮ⅡA对三阴性乳腺癌细胞MDA-MB-231迁移的抑制作用及其分子机制。方法:选取三阴性乳腺癌细胞MDA-MB-231,利用细胞增殖实验检测丹参酮ⅡA对MDA-MB-231细胞增殖的作用,并筛选适宜的药物浓度;应用划痕实验检测丹参酮ⅡA对MDA-MB-231细胞迁移率的影响;Western Blot法检测丹参酮ⅡA对G蛋白偶联雌激素受体(G protein-coupled estrogen receptor,GPER)及基质金属蛋白酶9(Matrix metalloprotein-9,MMP-9)表达的影响。结果:细胞增殖实验结果显示,丹参酮ⅡA可以抑制MDA-MB-231细胞增殖,且呈剂量依赖性(P<0.05);划痕实验结果显示,丹参酮ⅡA可以抑制MDA-MB-231细胞迁移,且呈剂量依赖性(P<0.01),加入GPER特异性抑制剂G15后迁移率有所上升(P<0.01)。Western Blot结果显示,丹参酮ⅡA可以显著下调GPER和MMP-9蛋白的表达水平并呈剂量依赖性(P<0.05),加入GPER特异性抑制剂G15后,MMP-9表达有所上升(P<0.01)。结论:丹参酮ⅡA可以抑制三阴性乳腺癌细胞MDA-MB-231迁移,其机制可能与抑制GPER介导的MMP-9表达相关。

Objective: To explore the anti-migration effect of Tanshinone ⅡA on triple negative breast cancer cells MDA-MB-231and its molecular mechanism based on in vitro assays. Methods: Triple negative breast cancer cells MDA-MB-231 were selected, and the effect of Tanshinone ⅡA on the proliferation of MDA-MB-231 cells was detected by cell proliferation assay. In addition, the optimal drug concentration was screened. The effect of Tanshinone ⅡA on the migration rate was detected by Wound-Healing assay. Western blot was performed to detect the expression of G protein-coupled estrogen receptor(GPER) and Matrix metalloprotein-9(MMP-9).Results: The results of cell proliferation assay showed that Tanshinone Ⅱ A could inhibit the proliferation of MDA-MB-231 cells in a dose-dependent manner(P<0.05). Wound-Healing assay showed that the Tanshinone Ⅱ A could inhibit the migration rate in a dose-dependent manner(P<0.01). Besides, the migration rate increased after administration of GPER-specific inhibitor, G15(P<0.01).Western Blot showed that Tanshinone ⅡA significantly downregulated expressions of GPER and MMP-9 proteins in a dose-dependent manner(P<0.05). The expression of MMP-9 increased after administration of GPER-specific inhibitor G15(P<0.01). Conclusion:Tanshinone ⅡA could inhibit the migration of triple-negative breast cancer cell MDA-MB-231, and the mechanism may be related to the inhibition of GPER-mediated MMP-9 expression.