炎性反应微环境对结肠癌细胞上皮向间质转化的作用  

Effects of inflammatory microenvironment on the epithelial to mesenchymal transition of colon cancer cells

作　　者：李袁飞[1] 赵和平[1] 朱国强 刘静 贾军梅[1] 张革红[1] 郭亚荣[1] 宋彬[1] 

机构地区：[1]山西医科大学第一医院肿瘤科,太原030001 [2]普外科 [3]解放军第二六四医院烧伤整形科

出　　处：《中华消化杂志》2014年第3期164-169,共6页Chinese Journal of Digestion

基　　金：基金项目：国家自然科学青年基金（81201956）;山西省自然科学基金（2013011043-5）;高校优秀科技创新基金（C01201013）

摘　　要：目的探讨炎性反应微环境中TNF-α和TGF-β1对结肠癌细胞上皮向间质转化的作用。方法以10#g／LTGF-β1或20μg／LTNF～α或两者混合后分别作用于人结肠癌细胞系SW480细胞、HCTll6细胞和经转化生长因子G受体2（TGFBR2）干扰的SW480细胞，单纯SW480细胞和HCTll6细胞分别设为空白对照，分别于作用24、48和72h观察各组细胞形态变化。采用实时定量RT-PCR和Western印迹法检测各组上皮钙黏素、神经钙黏素和波形蛋白的表达变化。通过细胞迁移实验检测TGFBR2干扰后细胞迁移能力的变化。采用单因素方差分析进行统计学处理。结果不同时间点，TGF-β1和TNF-α分别作用于sW480细胞、HCTll6细胞和经TGFBR2干扰的SW480细胞，其细胞形态均未发生明显的变化；TGF-β1和TNF-α混合作用于以上3组细胞，其中8W480细胞和经TGFBR2干扰的SW480细胞向成纤维细胞样形态变化。与TGF-β1和TNF-α混合作用的SW480组以及SW480空白组相比，经TGFBR2干扰的sw480组中上皮钙黏素mRNA表达（0．58±0．23比0．80±O．12比0．23±0．03）明显下调，神经钙黏素tuRNA表达（1．10±0．10比0．60±0．11比1．34±0．20）显著升高，差异均有统计学意义（F=46．92、47．93，P均〈0．05），而波形蛋白mRNA表达差异无统计学意义（P〉0．05）。TGF-β1和TNF-α混合作用的HCTll6组与HcTll6空白组相比，上皮钙黏素mRNA表达（0．22±0．03比0．92±0．14）明显下调，神经钙黏素（1．40±0．05比0．46±0．02）和波形蛋白（1．60±0．12比0．78±0．11）的mRNA表达均显著升高，差异均有统计学意义（F=72．59、23．67、45．19，P均〈0．05）。各组上皮钙黏素、神经钙黏素和波形蛋白的蛋白表达与mRNA表达一致。经TGFBR2干扰的SW480组A570为102．7±8．5，TGF-β1和TNF-α混合作用的SW480组和SW480空白组分别为20．1±7．6、18．3±11．2，差异有统计学意义（F-108．92，P〈0．05）。提示TGObjective To explore the effects of tumor necrosis factor （TNF-α） and transforming growth factor β1 （TGF-β1） in the inflammatory microenvironment on the epithelial to mesenchymal transition （EMT） of colon cancer cells. Methods SW480, HCT116 and SW480 interfered with transforming growth factor beta receptor 2 （TGFBR2） cells were treated with TGF-β1 （10μg/L）, TNF-α （20 μg/L）, or a combination of both. Untreated SW480 and HCT116 cells were set as blank controls. The morphologic changes of cells of each group were observed at 24, 48 and 72 hour. The expressions ofE-cadherin, N-eadherin and vimentin of each group were detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot. The cell migration ability of cells interfered with TGFBR2 was tested by cell migration assay. The data were analyzed by one way analysis of variance. Results There were no obvious morphologic changes in SW480 cells, HCTll6 cells and SW480 interfered with TGFBR2 at different time point. Among the three groups of cells treated with TNF-α and TGF-β1 combination, the morphologic changes to fibroblast were observed in the SW480 cells and SW480 interfered with TGFBR2. Compared with the group of SW480 cells treated with TNF-α and TGF-β1 and the group of SW480 blank controls, the expression of E-cadherin at mRNA level of the group of SW480 interfered with TGFBR2 significantly decreased （0. 58±0.23 vs 0. 80± 0. 12 vs 0. 23±0.03） and the expression of N-cadherin at mRNA level remarkably increased （1. 10±0. 10 vs 0. 60±0. 11 vs 1. 34±0.20）, the differences were statistically significant （F= 46. 92,47. 93, bofh P 〈0.05）. There was no statistically significant differences in the expression of vimintin （P〉0.05）. Compared with HCT116 blank control group, the expression of E-cadherin at mRNA level of HCTll6 treated with TNF-α and TGF-β1 combination significantly decreased （0. 92±0.14 vs 0. 22±0.03）, the expression of N-cadherin（0. 46±0.02 vs 1.40±0.05） an

关 键 词：结肠肿瘤 上皮向间质转化 转化生长因子Β1 肿瘤坏死因子Α 

分 类 号：R735.35[医药卫生—肿瘤]