NOX4在BEAS-2B细胞上皮间质转化过程中的作用
Role of NOX4 in epithelial-mesenchymal transition process of BEAS-2B cells

目的:探索非吞噬细胞氧化酶4(NOX4)在人肺正常上皮细胞BEAS-2B上皮间质转化过程中的作用。方法:选取对数生长期的BEAS-2B细胞,分为空白对照组、NOX4抑制剂二苯基氯化碘(DPI)组、TGF-β组和TGF-β+DPI组。采用Western blot检测E-cadherin、Vimentin、NOX4的表达,用划痕实验检测细胞迁移能力,流式细胞仪检测活性氧(ROS)水平。结果:TGF-β可增加BEAS-2B细胞NOX4、Vimentin的表达及ROS水平,降低E-cadherin的表达以及划痕宽度,DPI可以抑制NOX4、Vimentin的表达及ROS水平,增加E-cadherin的表达以及划痕宽度(P<0.001); DPI可拮抗TGF-β对NOX4、E-cadherin、ROS的作用(P<0.05)。结论:抑制NOX4的表达可以抑制TGF-β诱导的BEAS-2B细胞上皮间质转化,进而抑制BEAS-2B细胞的迁移能力。
Aim: To explore the role of non-phagocytic oxidase 4(NOX4)in the epithelial-mesenchymal transition(EMT)of BEAS-2B cells.Methods: BEAS-2B cells at logarithmic growth phase were allocated into 4 groups, blank control group, TGF-β group, DPI(an inhibitor of NOX4)group, and TGF-β+DPI group. Western blot was used to detect the expression of E-cadherin, Vimentin and NOX4. The cell migration ability was detected by scratch experiment. The reactive oxygen(ROS)was detected by flow cytometry.Results: It was shown that TGF-β could increase the expression of NOX4, Vimentin and ROS, and reduce the expression of E-cadherin and the width of scratches(P<0.001). DPI could inhibit the expression of NOX4, Vimentin and ROS, and increase the expression of E-cadherin and the width of scratches(P<0.001). The combination of DPI andTGF-β had antagonistic effects(P<0.05).Conclusion: Under the experimental conditions, inhibition of NOX4 expression can inhibit TGF-β-induced BEAS-2B cells' EMT process, thus inhibit the migration of BEAS-2B cells