Xue Teng, Xu Xiaoyan, Xia Yanqiu, Ren Ya'nan, Zhou Fang
Department of Occupational Health, College of Public Health, Zhengzhou University, Zhengzhou 450001, China.
Wei Sheng Yan Jiu. 2017 Jul;46(4):615-620.
To explore the role of NADPH oxidase 4( NOX4) in transforming growth factor-β( TGF-β)-induced A549 cells migration.
The A549 cells were allocated into five groups: TGF-β( stimulation) group, Normal control group, DPI( NOX4 inhibitor) group, TGF-β + DPI group, and DMSO( solvent control)group. The level of ROS were detected by flow cytometry instrument. The level of NOX4, snail and E-cadherin protein were detected by western blot. Use scratches experiment to express the change of A549 cells migration.
After the quantification by Quantity One software, the expression of NOX4 in TGF-β group is( 1. 80 ± 0. 07), the TGF-β +DPI group is( 0. 49 ± 0. 03)( F = 327. 071, P < 0. 001). The change of EMT related protein: the expression of snail protein in TGF-β group is( 9. 0 ± 0. 6), the TGF-β + DPI group is( 1. 8 ± 0. 3)( F = 119. 097, P < 0. 001), the expression of E-cadherin protein in TGF-β group is( 0. 5 ± 0. 1), the TGF-β + DPI group is( 3. 3 ± 0. 3)( F = 71. 063, P <0. 001). These aboveresult indicate that DPI can inhibit the expression of NOX4 and EMTprogress in A549 cells. Then TGF-β + DPI group compared with the TGF-β group, the scratch healing rate is decreased( F = 33. 899, P < 0. 001). It illustrates that DPI can inhibit migration ability of A549 cells.
After the NOX4 was inhibited by DPI, TGF-β-induced migration of A549 cells was inhibited. And this phenomenon is associated with the progress of TGF-β-induced EMT.
探讨NADPH氧化酶4（NOX4）在转化生长因子-β（TGF-β）诱导的A549细胞迁移中的作用。
将A549细胞分为五组：TGF-β（刺激）组、正常对照组、DPI（NOX4抑制剂）组、TGF-β + DPI组和二甲基亚砜（溶剂对照）组。采用流式细胞仪检测活性氧（ROS）水平。采用蛋白质印迹法检测NOX4、蜗牛蛋白和E-钙黏蛋白的蛋白水平。采用划痕实验观察A549细胞迁移的变化。
经Quantity One软件定量分析，TGF-β组中NOX4的表达为（1.80±0.07），TGF-β + DPI组为（0.49±0.03）（F = 327.071，P < 0.001）。上皮-间质转化（EMT）相关蛋白的变化：TGF-β组中蜗牛蛋白的表达为（9.0±0.6），TGF-β + DPI组为（1.8±0.3）（F = 119.097，P < 0.001），TGF-β组中E-钙黏蛋白的表达为（0.5±0.1），TGF-β + DPI组为（3.3±0.3）（F = 71.063，P < 0.001）。上述结果表明，DPI可抑制A549细胞中NOX4的表达及EMT进程。与TGF-β组相比，TGF-β + DPI组的划痕愈合率降低（F = 33.899，P < 0.001）。这说明DPI可抑制A549细胞的迁移能力。
DPI抑制NOX4后，TGF-β诱导的A549细胞迁移受到抑制。且此现象与TGF-β诱导的EMT进程相关。
