肾小管上皮-间质细胞转分化模型的建立

Establishment of a model of epithelial-mesenchymal transdifferentiation in HK-2 cells

目的探讨转化生长因子β1(TGF-β1)在人肾小管上皮细胞转分化中的作用,建立肾小管上皮-间质细胞转分化(TEMT)模型。方法实验分为正常对照组和TGF-β1处理组,利用TGF-β1处理人肾小管上皮细胞(HK-2)24 h,分别应用Western blot和免疫荧光方法检测上皮细胞和间质细胞分子标记的表达变化;应用划痕和Transwell assay方法检测细胞迁移及侵袭能力。结果与正常对照组比较,TGF-β1处理组HK-2细胞呈梭形,上皮细胞标记分子E-cadherin表达减弱,差异有统计学意义(P<0.01),间质细胞标记分子骨架蛋白Vimentin和β-catenin表达增强,差异有统计学意义(P<0.01);细胞的迁移及侵袭能力亦明显增强。结论 TGF-β1刺激后HK-2细胞促进上皮细胞转化为间质细胞。

Objective To investigate the roles of transforming growth factor β1（TGF-β1） （luring transdifferentiation of human renal tubular epithelial cells, and to establish a model of epithelial-mesenchymal transdifferentiation.Methods Tile experiment was divided into control and experimental groups. Human renal tubular epithelial cells （HK-2） were treated by TGF-β1 for 24 hours. The molecular markers in epithelial or mesenchymal eells were investigated by using Western blot and immune-fluorescence. To determine migration and invasion, scratching test and transwell assay were performed. Results The mesenchymal morphology was exhibited in HK-2 cells treated for 24 h by TGF-β1 （10 ng/ml）. Experiment data also showed that TGF-β1 can increase the expression of the mesenchymal phenotype markers （Vimentin, β-catenin）, and repress the expression of the epithelial phenotype marker （E-cadherin） in HK-2 cells. At the same time, compared with the control and the TGF-β1 groups, scratching test showed that TGF-β1 promoted HK-2 cell migration. The difference was statistically significant （P 〈 0.01 ）. In terms of invasion, a statistically significant difference was observed between the control group and TGF-β1 group （P 〈0. 01 ）. Conclusion Taken together, our findings provide the cell model of EMT in vitro using TGF-β1.