NF-κB对A549细胞上皮间质转化进程的影响

Effects of NF-κB on epithelial-mesenchymal transformation of A549 cells

目的:探讨NF-κB对A549细胞上皮间质转化(EMT)进程的影响。方法:选择处于对数生长期的A549细胞,分为空白组、TGF-β组、TGF-β+BAY11-7082(NF-κB抑制剂)组、BAY11-7082组和溶剂(二甲基亚砜)对照组。24 h后,采用Western blot法检测EMT相关蛋白(E-cadherin、Vimentin和Snail)以及NF-κB(p65)、NOX4蛋白的表达,采用DCFH-DA探针法检测ROS的水平,利用划痕实验检测A549细胞的迁移能力。结果:与空白组比较,TGF-β组细胞中Vimentin、Snail蛋白的表达水平升高,E-cadherin蛋白的表达水平降低,划痕宽度变窄,同时活化NF-κB(p65)并提高NOX4蛋白表达水平以及ROS的水平(P均<0.05);与TGF-β组比较,TGF-β+BAY11-7082组细胞中Vimentin、Snail蛋白的表达水平降低,E-cadherin蛋白的表达水平升高,划痕宽度变宽,同时NOX4蛋白表达水平以及ROS的水平降低(P均<0.05)。结论:抑制NF-κB可能通过降低NOX4的表达以及ROS的产生抑制TGF-β诱导的A549细胞的EMT进程,降低细胞的迁移能力。

Aim:To investigate the effects of NF-κB on epithelial mesenchymal transition(EMT)of A549 cells.Methods:A549 cells at logarithmic growth phase were allocated into blank group,TGF-βgroup,TGF-β+BAY11-7082(NF-κB inhibitor)group,BAY11-7082 group and DMSO(solvent control)group.After 24 hours,Western blot was performed to analyze the expressions of EMT-related proteins(E-cadherin,Vimentin,Snail),NF-κB(p65)and NOX4.DCFH-DA probe was used to detect the level of ROS,and scratch assay was used to detect cell migration ability.Result:Compared with blank group,in TGF-βgroup,the expression levels of Vimentin and Snail elevated,while that of E-cadherin reduced,the width of scratches decreased,NF-κB(p65)was activated,and the levels of NOX4 and ROS were up-regulated(P<0.05).Compared with TGF-βgroup,in TGF-β+BAY11-7082 group,the above changes were partly reversed(P<0.05).Conclusion:It is shown that inhibiting the expression of NF-κB might restrain TGF-β-induced A549 cells′EMT process by decreasing the levels of NOX4 and ROS,and then inhibit the migration of A549 cells.