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[Role of nicotinamide vadenine dinucleotide phosphate oxidase 4 in transforming growth factor-β-induced A549 cell
Teng Xue1, Xiaoyan Xu1, Yanqiu Xia1
1Department of Occupational Health, College of Public Health, Zhengzhou University, Zhengzhou 450001, China.
Objective:
To explore the role of NADPH oxidase 4( NOX4) in transforming growth factor-β( TGF-β)-induced A549 cells migration.
Methods:
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.
Results:
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.
Conclusion:
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.
Insights
Inhibiting NADPH oxidase 4 (NOX4) with DPI reduces transforming growth factor-beta (TGF-β)-induced migration in A549 cells. This effect is linked to the suppression of epithelial-mesenchymal transition (EMT).
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Transforming growth factor-beta (TGF-β) is a key regulator of cellular processes, including cell migration.
- NADPH oxidase 4 (NOX4) is implicated in various cellular functions and diseases.
- A549 cells are a human lung adenocarcinoma cell line commonly used in cancer research.
Purpose of the Study:
- To investigate the role of NOX4 in TGF-β-induced migration of A549 cells.
- To determine if inhibiting NOX4 affects TGF-β-induced epithelial-mesenchymal transition (EMT).
Main Methods:
- A549 cells were treated with TGF-β and/or the NOX4 inhibitor DPI.
- Reactive oxygen species (ROS) levels were measured by flow cytometry.
- Expression of NOX4, snail, and E-cadherin proteins was assessed using Western blot.
- Cell migration was evaluated using scratch healing assays.
Main Results:
- TGF-β stimulation increased NOX4 expression and promoted EMT, evidenced by increased snail and decreased E-cadherin.
- DPI treatment significantly reduced NOX4 expression and inhibited TGF-β-induced EMT.
- DPI treatment also decreased the scratch healing rate of A549 cells, indicating reduced migration ability.
Conclusions:
- NOX4 plays a crucial role in TGF-β-induced migration of A549 cells.
- Inhibition of NOX4 by DPI effectively suppresses TGF-β-induced cell migration.
- The observed inhibition of migration is associated with the suppression of TGF-β-induced EMT.
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