Nintedanib inhibits epithelial-mesenchymal transition in A549 alveolar epithelial cells through regulation of the TGF-β/Smad pathway.

Abstract

BACKGROUND:

Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic lung disorder. Recent studies have suggested that epithelial-mesenchymal transition (EMT) of alveolar epithelial cells influences development of pulmonary fibrosis, which is mediated by transforming growth factor β (TGF-β). Tumor necrosis factor α (TNF-α), an important proinflammatory cytokine in IPF, has been shown to enhance TGF-β-induced EMT. Nintedanib, a multiple tyrosine kinase inhibitor that is currently used to treat IPF, has been shown to suppress EMT in various cancer cell lines. However, the mechanism of EMT inhibition by nintedanib and its effect on TGF-β and TNF-α signaling pathways in alveolar epithelial cells have not been fully elucidated.

METHODS:

A549 alveolar epithelial cells were stimulated with TGF-β2 and TNF-α, and the effects of nintedanib on global gene expression were evaluated using microarray analysis. Furthermore, Smad2/3 phosphorylation was assessed using western blotting.

RESULTS:

We found that in A549 cells, TGF-β2 and TNF-α treatment induces EMT, which was inhibited by nintedanib. Gene ontology analysis showed that nintedanib significantly attenuates the gene expression of EMT-related cellular pathways and the TGF-β signaling pathway, but not in the TNF-α-mediated signaling pathway. Furthermore, hierarchical cluster analysis revealed that EMT-related genes were attenuated in nintedanib-treated cells. Additionally, nintedanib was found to markedly suppress phosphorylation of Smad2/3.

CONCLUSION:

Nintedanib inhibits EMT by mediating EMT-related gene expression and the TGF-β/Smad pathway in A549 alveolar epithelial cells.

• Authors: Hiroaki Ihara, Yoichiro Mitsuishi, Motoyasu Kato, Fumiyuki Takahashi, Ken Tajima, Takuo Hayashi, Moulid Hidayat, Wira Winardi, Aditya Wirawan, Daisuke Hayakawa, Koichiro Kanamori, Naohisa Matsumoto, Toshifumi Yae, Tadashi Sato, Shinichi Sasaki, Kazuya Takamochi, Yoshiyuki Suehara, Dai Ogura, Shin-Ichiro Niwa, Kenji Suzuki, Kazuhisa Takahashi
• Journal: Respiratory investigation (Respir Investig) Vol. 58 Issue 4 Pg. 275-284 (Jul 2020) ISSN: 2212-5353 [Electronic] Netherlands
• PMID: 32359980 (Publication Type: Journal Article)
• Copyright: Copyright © 2020 The Japanese Respiratory Society. Published by Elsevier B.V. All rights reserved.

Chemical References

• Indoles
• SMAD2 protein, human
• Smad2 Protein
• Transforming Growth Factor beta2
• Tumor Necrosis Factor-alpha
• nintedanib

Topics

• A549 Cells
• Epithelial Cells (metabolism)
• Epithelial-Mesenchymal Transition (drug effects, genetics)
• Gene Expression (drug effects)
• Humans
• Indoles (pharmacology)
• Phosphorylation (drug effects)
• Pulmonary Alveoli (cytology, physiology)
• Signal Transduction (drug effects, genetics)
• Smad2 Protein (metabolism)
• Transforming Growth Factor beta2 (metabolism, pharmacology)
• Tumor Necrosis Factor-alpha (pharmacology)