TGF-β1促进食管癌上皮间质转化的发生发展

TGF-β1 promotes esophageal cancer Epithelial-Mesenchymal-Transition process

目的探讨转化生长因子(TGF-β1)在促进食管癌Eca109细胞增殖、迁移、侵袭和上皮间质转化中的作用。方法利用TGF-β1受体抑制剂和病毒转染TGF-β1干扰RNA(siRNA)处理Eca109细胞,分为3组:实验组(病毒稳定TGF-β1 siRNA转染Eca109细胞)、阴性对照组(转染无关序列Eca109细胞)、空白对照组(正常未经任何处理的Eca109细胞),采用荧光定量PCR(qRT-PCR)技术检测TGF-β1及上皮间质转化(EMT)相关指标E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、锌指转录因子(Snail)]在mRNA水平表达情况;EMT相关指标蛋白(Ecadherin、Vimentin)的表达;细胞增殖实验(MTT)、细胞划痕实验及Transwell法分别检测细胞增殖、迁移、侵袭的变化情况。结果通过TGF-β1受体抑制剂(0、0.1、1、10μm/m L)处理Eca109细胞后,在mRNA水平TGF-β1表达逐渐降低,E-cadherin表达逐渐升高,Vimentin、Snail表达逐渐降低(P<0.05);在蛋白水平,E-cadherin表达逐渐升高,Vimentin表达逐渐降低,细胞增殖和迁移能力也逐渐降低(P<0.05)。实验组与空白对照组和阴性对照组相比,在mRNA水平实验组TGF-β1表达降低(P<0.05);在蛋白水平,实验组E-cadherin表达增高,Vimentin表达降低;实验组细胞增殖、迁移和侵袭能力均降低(P<0.05)。结论在细胞水平,TGF-β1可促进食管癌细胞的增殖和上皮间质转化的进程。

Objective To investigate whether TGF-β1 could promote cell proliferation,migration,invasion and epithelial-mesenchymal-transition（ EMT） process in Eca109. Methods With stimulation of TGF-β1 receptor inhibitor,the expression of TGF-β1 and EMT-associated mRNAs were detected by qRT-PCR; the expression of EMTassociated proteins were detected by western blot. Then TGF-β1 siRNA was transfected into Eca109 cells by lentiviral vector,the expression level of TGF-β1 was detected by qRT-PCR,EMT-associated proteins were measured by western blot after transfection. MTT,Wound-healing assay and transwell assay were performed to detect cell proliferation,migration and invasion ability. Results Treatment of TGF-β1 receptor inhibitor,TGF-β1 and Vimentin,Snail mRNA were significantly decreased,while the expression of E-cadherin mRNA was increased（ P〈0. 05）.EMT-associated proteins could induce the down-regulation of Vimentin expression and up-regulate the E-cadherin expression. After transfected with TGF-β1 siRNA,the mRNA expression of TGF-β1 and the protein expression of Vimentin were down-regulated（ P〈0. 05）,while the protein expression of E-cadherin was up-regulated. Furthermore,treatment of TGF-β1 receptor inhibitor or TGF-β1 siRNA inhibited cell migration,proliferation and invasion ability. Conclusion These data suggest that TGF-β1 promotes esophageal cancer epithelial-mesenchymal-transition process.