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目的探究流体剪切力(fluid shear stress,FSS)对喉鳞癌Hep2细胞发生上皮-间充质转化(epithelial mesenchymal transition,EMT)的影响。方法对Hep2细胞加载140 m Pa的FSS,观察不同时间点细胞的形态学变化;划痕实验检测力学加载后Hep2细胞迁移能力的变化;共聚焦显微镜下观察细胞骨架蛋白F-actin的分布;Western blotting检测EMT相关蛋白的表达。结果 FSS力学加载后,Hep2细胞形态由多边形向梭形转变,撤销FSS后,细胞恢复初始的多边形;Hep2细胞迁移能力的增加依赖于FSS加载时间。FSS促使细胞骨架蛋白F-actin重排,从而增强Hep2细胞的迁移行为。FSS使Hep2细胞EMT标志蛋白发生了时序性变化。结论 FSS是诱导Hep2细胞发生EMT的一个重要物理因素。
Objective To investigate the effect of fluid shear stress (FSS) on epithelial mesenchymal transition (EMT) in Hep2 cells. Methods Hep2 cells were loaded with 140 mPa FSS, and the morphological changes of cells were observed at different time points. Scratch assay was used to detect the change of Hep2 cell migration after mechanical loading. The distribution of F-actin was observed under confocal microscopy. blotting was used to detect the expression of EMT related proteins. Results After FSS loading, the morphology of Hep2 cells changed from polygons to fusions. After FSS was withdrawn, the cells recovered to the original polygons; the increase of Hep2 cell migration ability was dependent on the FSS loading time. FSS promotes cytoskeletal protein F-actin rearrangement, thereby enhancing Hep2 cell migration behavior. FSS caused the temporal change of EMT marker protein in Hep2 cells. Conclusion FSS is an important physical factor for inducing EMT in Hep2 cells.