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Transforming Growth Factor-β1 (TGF-β1) Driven Epithelial to Mesenchymal Transition (EMT) is Accentuated by Tumour Necrosis Factor α (TNFα) via Crosstalk Between the SMAD and NF-κB Pathways

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Cancer Microenvironment

Abstract

Epithelial to mesenchymal transition (EMT) is a process by which an epithelial cell alters its phenotype to that of a mesenchymal cell and plays a critical role in embryonic development, tumour invasion and metastasis and tissue fibrosis. Transforming growth factor-β1 (TGF-β1) continues to be regarded as the key growth factor involved in driving EMT however recently tumour necrosis factor α (TNFα) has been demonstrated to accentuate TGF-β1 driven EMT. In this study we investigate how various signalling pathways contribute to this accentuated effect. A549 cells were treated with TGF-β1 (10 ng/ml), TNFα (20 ng/ml) or a combination of both for 72 h and EMT assessed. The effect of selective inhibition of the SMAD, MAPK and NF-κB pathways on EMT was assessed. A549 cells treated with TGF-β1 downregulate the expression of epithelial markers, increase the expression of mesenchymal markers, secrete matrix-metalloproteinases and become invasive. Significantly, TGF-β1 driven EMT is accentuated by co-treatment with TNFα. SMAD 3 inhibition attenuated TGF-β1 driven EMT but has no effect on the accentuation effect of TNFα. However, inhibiting IKKβ blocked both TGF-β1 driven EMT and the accentuating action of TNFα. Inhibiting p38 and ERK signalling had no effect on EMT. TNFα accentuates TGF-β1 driven EMT in A549 cells via a SMAD 2/3 independent mechanism involving the NF-κB pathway independent of p38 and ERK 1/2 activation.

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Acknowledgments

This work was supported by a research grant from the Medical Research Council UK (G0700861). LAB is supported by a Marie Curie fellowship award. AJF is supported by a GlaxoSmithKline clinical fellowship award. ADS is supported by a HEFCE Senior Lectureship. DAM is supported by the Wellcome Trust (WT086755MA).

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Authors and Affiliations

  1. Tissue Fibrosis and Repair Group, Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, UK, NE2 4HH

    Lee A. Borthwick, Aaron Gardner, Anthony De Soyza, Derek A. Mann & Andrew J. Fisher

  2. Department of Respiratory Medicine, Freeman Hospital, High Heaton, Newcastle upon Tyne, UK, NE7 7DN

    Anthony De Soyza & Andrew J. Fisher

  3. Institute of Cellular Medicine, Medical School, Newcastle University, Newcastle upon Tyne, UK, NE2 4HH

    Andrew J. Fisher

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  1. Lee A. Borthwick
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Correspondence to Andrew J. Fisher.

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Borthwick, L.A., Gardner, A., De Soyza, A. et al. Transforming Growth Factor-β1 (TGF-β1) Driven Epithelial to Mesenchymal Transition (EMT) is Accentuated by Tumour Necrosis Factor α (TNFα) via Crosstalk Between the SMAD and NF-κB Pathways. Cancer Microenvironment 5, 45–57 (2012). https://doi.org/10.1007/s12307-011-0080-9

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