Rho信号通路抑制剂Y27632抑制TGF-β诱导的MCF-7细胞上皮间质转化

摘要

许多研究表明在乳腺癌的发生过程中上皮间质转化(epithelial-to-mesenchymal transition，EMT)起到十分重要的作用，转化生长因子–β(transforming growth factor-β，TGF-β)是公认的可以引起EMT的重要因子，在EMT过程中涉及许多信号通路，但是Rho信号通路在其中的作用尚未报道。本文首先建立TGF-β诱导人乳腺癌细胞MCF-7上皮间质转化的模型，利用划痕实验检测细胞迁移能力，利用实时定量荧光PCR(real-time PCR)和免疫印迹(Western blot)实验检测间质标志基因的表达情况。进一步通过加入Rho通路抑制剂Y27632研究Rho通路在TGF-β诱导的MCF-7细胞EMT中的作用，结果证实TGF-β可以诱导MCF-7细胞EMT，促进MCF-7细胞迁移，而Y27632抑制TGF-β诱导的MCF-7细胞EMT过程及其迁移。%It is reported that epithelial-mesenchymal transition(EMT)plays an important role in the process of breast cancer development. TGF-βis reported to be the key factor during the EMT progress. Multiple signaling pathways are involved in EMT. Whether Rho/ROCK pathway is involved in EMT is unclear. First,TGF-β-induced EMT model was established. The migration abilities of MCF-7 were evaluated by wound-healing assay and the expression of mesenchymal markers were de-tected by real-time PCR and western blotting. To investigate the role of Rho pathway in TGF-β-induced MCF-7 cell EMT, specific inhibitor Y27632 was added in MCF-7 cells with/without TGF-β. The results showed that TGF-βdid induce EMT and migration,and Y27632 really inhibited TGF-β-induced EMT and migration in MCF-7.