去氢骆驼蓬碱对MCF7细胞增殖与迁移侵袭的影响

Effects of Harmine on the Proliferation,Migration and Invasion of Human Breast Cancer MCF7 Cells

目的:观察去氢骆驼蓬碱对人乳腺癌MCF7细胞增殖、迁移与侵袭能力的影响,并探究其潜在的分子机制。方法:体外常规培养人乳腺癌MCF7细胞,用不同浓度的去氢骆驼蓬碱(0、2.5、5、10、20、40、80μmol/L)处理细胞,采用MTT法测定其对细胞增殖能力的影响;依据MTT实验结果,确定后续实验组药物作用浓度(0、5、10、20μmol/L),采用克隆形成实验、细胞划痕实验、Transwell实验观察去氢骆驼蓬碱对细胞克隆形成、迁移和侵袭能力的影响;通过Western blot检测N-Cadherin、Vimentin及TGF-β通路相关蛋白的表达情况。结果:与对照组比较,在浓度为5、10、20μmol/L时,去氢骆驼蓬碱可显著抑制MCF7细胞的增殖(P<0.05),其作用呈浓度及时间依赖性;并能使MCF7细胞克隆形成数、细胞迁移率、细胞侵袭率均显著降低(P<0.05);Western blot结果显示,N-Cadherin、Vimentin、Stat3、TGF-β、Smad2、Smad3、Smad4等蛋白表达均显著下调(P<0.05)。结论:综上所述,去氢骆驼蓬碱可能介导TGF-β通路,抑制相关蛋白的表达,经间充质转换的调节作用,抑制MCF7细胞的增殖、迁移与侵袭能力。

Objective:To study the effects of harmine on the proliferation,migration and invasion of human breast cancer MCF7 cells,and to explore its potential molecular mechanism.Methods:Human breast cancer MCF7 cells were cultured in vitro and treated with different concentrations of harmine(0,2.5,5,10,20,40,80 μmol/L).MTT assay was used to determine the effect on cell proliferation.According to the results of the MTT assay,the appropriate concentrations of harmine for the following experimental group(0,5,10,20 μmol/L)were decided.The experiments of colony formation,healing wound and Transwell were used to observe the effects of harmine on cell clone formation,migration and invasion.And the Western blot was used to test the expressions of N-Cadherin,Vimentin and the related protein of TGF-β pathway.Results:Compared with the control group,the proliferation of MCF7 cells was inhibited significantly at the concentrations of 5,10,20 μmol/L(P<0.05),and in a concentration-time dependent,the number of clone formation,the ratio of healing wound and the invasion rate of MCF7 cells were significantly decreased(P<0.05).The results of Western blot showed that harmine could down-regulated the expressions of N-Cadherin,Vimentin,Stat3,TGF-β,Smad2,Smad3 and Smad4(P<0.05).Conclusion:In Conclusion,harmine may mediate TGF-β pathway,down-regulated the expressions of the related proteins,inhibit the proliferation,migration and invasion of MCF7 cells through the regulation of mesenchymal transformation.