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The RET/PTC3 oncogene activates classical NF-κB by stabilizing NIK

Oncogene volume 30, pages 87–96 (2011)Cite this article

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Abstract

The oncogenic fusion protein RET/PTC3 (RP3) that is expressed in papillary thyroid carcinoma (PTC) and thyroid epithelia in Hashimoto's thyroiditis activates nuclear factor-kappa B (NF-κB) and induces pro-inflammatory gene expression; however, the mechanism of this activation is unknown. To address this, we expressed RP3 in murine embryonic fibroblasts (MEFs) lacking key classical and noncanonical NF-κB signaling components. In wild-type MEFs, RP3 upregulated CCL2, CXCL1, granulocyte-macrophage colony-stimulating factor and tumor necrosis factor expression and activated classical but not noncanonical NF-κB. RP3-activated NF-κB in IκB kinase (IKK)β−/− MEFs but not IKKα- or NF-κB essential modulator (NEMO)-deficient cells and activation was inhibited by a peptide that blocks NEMO binding to the IKKs. RP3 increased the levels of NF-κB-inducing kinase (NIK) and did not activate NF-κB in NIK-deficient MEFs. Notably, NIK stabilization was not accompanied by TRAF3 degradation demonstrating that RP3 disrupts normal basal NIK regulation. Dominant-negative NIK blocked RP3-induced NF-κB activation and an RP3 signaling mutant (RP3Y588F) did not stabilize NIK. Finally, examination of PTC specimens revealed strong positive staining for NIK. We therefore conclude that RP3 activates classical NF-κB via NIK, NEMO and IKKα. Importantly, our findings reveal a novel mechanism for oncogene-induced NF-κB activation via stabilization of NIK.

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Acknowledgements

This work was supported by grants from the National Institutes of Health (RO1HL080612, R21CA135602 and T32CA009683, T32GM07229) and the Mari Lowe Center for Comparative Oncology.

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Authors and Affiliations

  1. Department of Animal Biology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA, USA

    R J Neely, C M Gray, K A McCorkell & M J May

  2. Department of Microbiology and Immunology/Otolaryngology,

    R J Neely & J L Rothstein

  3. Head and Neck Surgery, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA

    R J Neely & J L Rothstein

  4. Department of Otorhinolaryngology,

    M S Brose, J M Leibowitz & C Ma

  5. Head and Neck Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA

    M S Brose, J M Leibowitz & C Ma

  6. Inflammation Research, Amgen, Inc., Seattle, Washington, WA, USA

    J L Rothstein

  7. The Mari Lowe Center for Comparative Oncology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA, USA

    M J May

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  1. R J Neely
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Correspondence to M J May.

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Neely, R., Brose, M., Gray, C. et al. The RET/PTC3 oncogene activates classical NF-κB by stabilizing NIK. Oncogene 30, 87–96 (2011). https://doi.org/10.1038/onc.2010.396

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