【摘要】 目的:探讨流体切应力在肝癌细胞迁移中的作用。方法:利用平行平板流动腔对培养的肝癌HepG2细胞施加1.4dyn·(cm2)-1稳定层流切应力,作用2h、4h、8h等不同时间后,采用划痕法观察细胞迁移的变化,同时以免疫荧光法观察细胞骨架F-actin变化,采用Western blot测定相关蛋白的表达,采用透射电镜观察细胞间紧密连接的变化。在此基础上,分析在流体切应力作用下,肝癌细胞上皮标志物(E-Cadherin)和间充质标志物(N-Cadherin、Vimentin、Twist等)的表达和分布,及其对细胞紧密连接的影响。结果:流体切应力可促进HepG2细胞迁移,且呈时间依赖性(P<0.05);随着流体切应力加载时间的延长,肝癌HepG2细胞E-Cadherin表达下调,而N-Cadherin、Vimentin、Twist表达上调(P<0.05)。同时,HepG2细胞间紧密连接被打开;当撤除流体切应力,上述诱导作用则发生不同程度的逆转(P<0.05)。结论:流体切应力通过诱导肝癌细胞上皮-间充质转化引起细胞间紧密连接降低,进而促进细胞迁移。更多还原

【Abstract】 Objective:To investigate the effect of fluid shear stress on the migration of HepG2 cells.Methods:A parallel flow chamber was used to provide 1.4 dyn·(cm2)-1 steady laminar shear stress to HepG2 cells for 2 h,4 hand 8 h.Cell migration was evaluated by scratch wound assay;cytoskeleton F-actin was observed by immunofluorescence staining;the expression of related proteins was determined by Western blot;and the tight junctions between cells were observed by transmission electron microscopy(TEM).In addition,the expression and distribution of epithelial(E-cadherin)and mesenchymal markers(N-cadherin,Vimentin and Twist)in HepG2 cells were analyzed.Results:Fluid shear stress promoted the migration of HepG2 cells in a time-dependent manner.The expression of N-cadherin was down-regulated while the expression of N-cadherin,Vimentin and Twist was up-regulated in HepG2 cells with the durations of fluid shear stress(P<0.05).Meanwhile,the tight junction distance between cells was increased.When shear stress was withdrawn,the above effects were reversed in different degrees(P<0.05).Conclusion:Fluid shear stress induces epithelial-mesenchymal transition in HepG2 cells,which leads to decreased tight junctions between cells and promotes cell migration.更多还原