Smad4在TGF—β诱导人胆管癌细胞上皮-间质转化中的作用  

The role of Smad4 in TGF-β induced epithelial- mesenchymal transition（EMT） of human cholan- giocarcinoma cells

作　　者：林有智[1] 陈孝平[1] 陆玉蕾[2] 梁慧芳[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝脏外科,武汉430030 [2]广西医科大学附属肿瘤医院

出　　处：《临床外科杂志》2013年第5期340-342,共3页Journal of Clinical Surgery

基　　金：华中科技大学自主创新基金（中央高校基本业务费资助）项目（2011JC065）

摘　　要：目的 探讨Smad4在TGF-β诱导人胆管癌细胞上皮-间质转化（epithelial-mesenchymal transition,EMT）中的作用.方法 用转化生长因子β1（transforming growth factor-β1,TGF-β1）处理RBE人胆管癌细胞系24h后,观察其形态变化,采用Western Blotting检测TGF-β1处理0h、12h、24h、48h后RBE细胞N-钙黏蛋白（N-cadherin）和E-钙黏蛋白（E-cadherin）的表达.通过RNA干扰技术建立稳定干扰Smad4表达的细胞系,在无TGF-β1处理或TGF-β1处理24h后,观察Smad4干扰组与野生型组和空载体组细胞的形态差异,Western Blotting检测各组N-cadherin和E-cadherin的表达水平.结果 TGF-β1诱导RBE细胞的形态由上皮样向间质样转变,促进 N-cadherin表达增加和E-cadherin表达降低.在无TGF-β1处理时,Smad4干扰组细胞比野生型组和空载体组细胞的形态更趋于上皮样,E-cadherin表达水平也显著高于野生型组和空载体组（P＜0.05）.在TGF-β1处理24h后,Smad4干扰组细胞形态的间质样变化程度、N-cadherin表达量明显低于野生型组和空载体组（P＜0.05）;野生型组和空载体组细胞E-cadherin的表达缺失,而Smad4干扰组细胞仍表达较高水平的E-cadherin,两者间差异显著（P＜0.05）.结论 TGF-β依赖Smad4介导人胆管癌细胞发生上皮-间质转化.Objective To Investigate the role of Smad4 in TGF-βindueed epithelial-mesenehyreal transition（EMT） of human cholangioeareinoma cells. Methods Human eholangiocaxeinoma cell line RBE was treated with exogenous TGF-β1 ,followed by observation of morphological ehanges under inverted microscopy. Meanwhile,the expression of N-eadherin and E-eadherin was analyzed by Western Blotting. Stable Smad4 knockdown RBE cells were generated by RNA interference. In the presence or absence of TGF-β1, differences in cell morphology and expression of N-eadherin and E-eadherin were evaluated between Smad4 knockdown group and wild-type and vector control group by light microscope and Western Blotting. Results TGF-β1 induced the switch from epithelioid morphology to mesenehymal morphology in RBE cells, and led to increased expression of N-eadherin and decreased expression of E-eadherin, eompared with TGF-β1-untreated eells. In the absence of TGF-β1, Smad4 knockdown eells showed more like epithelioids and higher expression level of E-cadherin（ P 〈 0.05 ）than wild-type and vector control eells. When treated with TGF-[31, Smad4 knockdown eells were less prone to mesenehymal morphology accompa- nied with a lower expression level of N-eadherin, eornpared with wild-type and veetor control cells （ P 〈 0.05 ）. Whereas wild-type and vector eontrol cells lost the expression of E-eadherin, Smad4 knoekdown cells still showed a considerable expression level of E-eadherin after TGF-[31 treatment（ P 〈 0.05 ）. Conlusion TGF-β induces EMT of RBE human eholangioeareinoma cells dependent on Smad4.

关 键 词：SMAD4 转化生长因子Β 上皮-间质转化 胆管癌 

分 类 号：R735.8[医药卫生—肿瘤]