TGF-β promote epithelial-mesenchymal transition via NF-κB/NOX4/ROS signal pathway in lung cancer cells

Epithelial-mesenchymal transition (EMT), transforming growth factor beta(TGF-beta) and reactive oxygen species(ROS) plays a central role in cancer metastasis. Moreover, nicotinamide adenine dinucleotide phosphate 4(NOX4) is one of the main sources of ROS in lung cancer cells suggesting that NOX4 is associated with tumor cell migration. NF-kappa B(Nuclear factor-Kappa-B) is known to regulate ROS-mediated EMT process by activating Snail transcription factor in A549 cells. The purpose of this study was to explore the relationship between NF-kappa B and NOX4 in ROS production during TGF-beta induced EMT process. Several fractions have been pooled to evaluates the EMT process on lung cancer cells through real-time PCR, Western Blot and flow cytometry with DCFH-DA probe etc. Cells proliferation and migration activities were monitored by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and wound healing assay respectively. The result showed that TGF-beta induction decreased the expression of E-cadherin, increased the Vimentin and the EMT transcription factor Snail in A549 cells. DPI (Diphenyleneiodonium chloride, an inhibitor of NOX4) inhibited the NOX4 expression and reduced ROS production induced by TGF-beta, but didn't affect the activation of NF-kappa B induced by TGF-beta (P > 0.05). BAY11-7082 (an inhibitor of NF-kappa B) inhibited the NF-kappa B (p65) expression and prevented the increase of NOX4 expression and ROS production induced by TGF-beta (P < 0.001), which has also verified reduced TGF-beta induced cell migration by inhibiting the EMT process, and also reduced cell proliferation of A549 cells (P < 0.001). The current research confirmed the TGF-beta mediated EMT process via NF-kappa B/NOX4/ROS signaling pathway, NF-kappa B and NOX4 are likely to be the potential therapeutic targets for lung cancer metastasis.

Automated summary

The study confirmed the TGF-beta mediated EMT process through the NF-kappa B/NOX4/ROS signaling pathway, suggesting NF-kappa B and NOX4 are potential therapeutic targets for lung cancer metastasis.