镰形棘豆总黄酮通过TGF-β/Smad3通路抑制骨肉瘤MG63细胞EMT

Inhibitory effect of total flavonoids from Oxytropis falciformis on EMT of osteosarcoma MG63 cells via TGF-beta/Smad3 pathway

目的基于TGF-β/Smad3通路探讨镰形棘豆总黄酮(FOF)抑制骨肉瘤MG63细胞上皮细胞间质转化(EMT)的分子机制。方法 MTT实验检测细胞增殖抑制情况;Transwell小室迁移实验观察细胞迁移能力变化;ELISA实验检测细胞分泌的TGF-β1水平;qPCR实验检测细胞中Snail1、Twist mRNA表达水平变化;Western blot实验检测细胞中E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、p-Smad3、磷酸化转化生长因子β受体Ⅱ(p-TGF-βRⅡ)蛋白表达水平变化。结果 FOF以时间和浓度依赖的方式抑制MG63细胞增殖,降低MG63细胞迁移及分泌TGF-β1的能力;并导致MG63细胞内Vimentin、p-Smad3、p-TGF-βRⅡ蛋白和Snail1、Twist mRNA表达水平显著减少(均P<0.05),而E-cadherin蛋白表达水平增加(P<0.05)。结论FOF对MG63细胞增殖具有抑制作用,并可通过抑制TGF-β1/Smads过度激活,逆转MG63细胞EMT,从而降低MG63细胞转移能力。

Objective To investigate the molecular mechanism of flavonoids of Oxytropis falcate(FOF)inhibiting epithelial-mesenchymal transition(EMT)in osteosarcoma MG63 cells based on the TGF-β/Smad3 pathway.Methods MTT assay was used to detect cell proliferation inhibition;Transwell chamber migration assay was used to observe cell migration ability;ELISA was used to detect the level of TGF-β1 secreted by cells;qPCR was used to detect the expression of Snail1 and Twist mRNA in cells.Results FOF inhibited the proliferation of MG63 cells in a time-and concentration-dependent manner,reduced the migration of MG63 cells and the ability to secrete TGF-β1,and resulted in a significant decrease in the expression levels of Vimentin,p-Smad3,p-TGF-βRⅡprotein and Snail1,Twist mRNA in MG63 cells,and an increase in E-cadherin protein.Conclusion FOF can inhibit the proliferation of MG63 cells and reverse the EMT of MG63 cells by inhibiting the over-activation of TGF-β1/Smads,thus reducing the metastasis ability of MG63 cells.