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TGF-β and NF-κB signal pathway cross-talk is mediated through TAK1 and SMAD7 in a subset of head and neck cancers

Oncogene volume 32, pages 1549–1559 (2013)Cite this article

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Abstract

Transforming growth factor-beta (TGF-β) has a dual role in epithelial malignancies, including head and neck squamous cell carcinoma (HNSCC). Attenuation of canonical TGF-β signaling enhances de novo tumor development, whereas TGF-β overexpression and signaling paradoxically promotes malignant progression. We recently observed that TGF-β-induced growth arrest response is attenuated, in association with aberrant activation of nuclear factor-κB (NF-κB), a transcription factor, which promotes malignant progression in HNSCC. However, what role cross-talk between components of the TGF-β and NF-κB pathways plays in altered activation of these pathways has not been established. Here, we show TGF-β receptor II and TGF-β-activated kinase 1 (TAK1) are predominantly expressed in a subset of HNSCC tumors with nuclear activation of NF-κB family member RELA (p65). Further, TGF-β1 treatment induced sequential phosphorylation of TAK1, IKK, IκBα and RELA in human HNSCC lines. TAK1 enhances TGF-β-induced NF-κB activation, as TAK1 siRNA knockdown decreased TGF-β1-induced phosphorylation of IKK, IκB and RELA, degradation of IκBα, RELA nuclear translocation and DNA binding, and NF-κB-induced reporter and target gene transcription. Functionally, TAK1 siRNA inhibited cell proliferation, migration and invasion. Celastrol, a TAK1 inhibitor and anti-inflammatory compound used in traditional Chinese medicine, also decreased TGF-β1-induced phosphorylation of TAK1 and RELA, and suppressed basal, TGF-β1- and tumor necrosis factor-alpha (TNF-α)-induced NF-κB reporter gene activity. Celastrol also inhibited cell proliferation, while increasing sub-G0 DNA fragmentation and Annexin V markers of apoptosis. Furthermore, TGF-β and RELA activation promoted SMAD7 expression. In turn, SMAD7 preferentially suppressed TGF-β-induced SMAD and NF-κB reporters when compared with constitutive or TNF-α-induced NF-κB reporter gene activation. Thus, cross-talk by TGF-β via TAK1 and NF-κB promotes the malignant phenotype of HNSCC. Moreover, NF-κB may contribute to the downstream attenuation of canonical TGF-β signaling through increased SMAD7 expression. Celastrol highlights the therapeutic potential of agents targeting TAK1 as a key node in this pro-oncogenic TGF-β-NF-κB signal pathway.

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Acknowledgements

Supported by intramural projects ZIA-DC-000016 and ZIA-DC-000073 from the National Institute on Deafness and Other Communication Disorders, NIH. Dr Contag Wise was supported by the Clinical Research Training Program, a public–private partnership supported jointly by the NIH and Pfizer Inc. The authors wish to thank Dr Ning T Yeh (NIDCD/NIH) and Chris Silvin (NHGRI/NIH) for their technical assistance. We would like to thank Drs Adam Glick and Ashok Kulkarni for review of the manuscript and helpful comments.

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Authors and Affiliations

  1. Tumor Biology Section, Head and Neck Surgery Branch, National Institute on Deafness and Other Communication Disorders, NIH, Bethesda, MD, USA

    C Freudlsperger, Y Bian, S Contag Wise, J Burnett, J Coupar, X Yang, Z Chen & C Van Waes

  2. Department of Oral and Maxillofacial Surgery, University Hospital Tuebingen, Tuebingen, Germany

    C Freudlsperger

  3. Clinical Research Training Program, a public–private partnership supported jointly by the NIH and Pfizer Inc,

    S Contag Wise

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  1. C Freudlsperger
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Freudlsperger, C., Bian, Y., Contag Wise, S. et al. TGF-β and NF-κB signal pathway cross-talk is mediated through TAK1 and SMAD7 in a subset of head and neck cancers. Oncogene 32, 1549–1559 (2013). https://doi.org/10.1038/onc.2012.171

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