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Simultaneous downregulation of CDK inhibitors p18Ink4c and p27Kip1 is required for MEN2A-RET-mediated mitogenesis

Oncogene volume 26, pages 554–570 (2007)Cite this article

Abstract

Multiple endocrine neoplasia type 2A (MEN2A) is predisposed by mutations in the RET proto-oncogene. Low expression of the cyclin-dependent kinase inhibitor (CDKI) p27Kip1 is present in thyroid tumors, and recent evidence demonstrates p27 downregulation by the active RET mutant, RET/PTC1, found in papillary thyroid carcinoma. This implicates decreased p27 activity as an important event during thyroid tumorigenesis. However, p27−/− mice develop MEN-like tumors only in combination with loss of another CDKI, p18Ink4c. This suggests that p18 and p27 functionally collaborate in suppression of tumorigenesis, that loss of both is critical in the development of MEN tumors and that both p18 and p27 are regulated by RET. We report that induction of the constitutively active MEN2A-specific RET mutant, RET2AC634R, correlates with reduced p18/p27, and elevated cyclin D protein levels, leading to increased CDK activity, increased pRb phosphorylation and proliferation under growth arrest conditions. Mechanistically, RET2A represses p18/p27 mRNA levels while elevating cyclin D1 mRNA levels. RET2A expression also correlates with decreased p27 protein stability. RET2A-mediated regulation of p18 and p27, but not of cyclins D1 and D2, requires functional mitogen-activated protein kinase signaling. Additionally, RET2A-dependent p18 repression is required and sufficient to increase cell proliferation. Perhaps most significantly, MEN2A adrenal tumors also display these changes in cell cycle expression profile, demonstrating the biological relevance of our cell culture studies. Our results demonstrate for the first time that RET2A regulates p18, and suggest that loss of not only p27 but also of p18 expression is a key step in MEN tumorigenesis.

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Acknowledgements

We thank Cheryl Holdman, Kathy Ragheb, Dr Joseph Ogas and Dr Richard Kuhn for technical assistance and several colleagues for critical review of the manuscript. This work was supported by the American Cancer Society (RSG-03-236-01-MGO), Purdue University Department of Biological Sciences and the Purdue University Cancer Center. We thank AH Bruggink, Coordinator University Medical Center Utrecht-Biobank, for providing the normal human adrenal tissues and MEN2A tumor samples. We thank Michael Toole for his assistance in immunostaining of adrenal sections and quantitation of the positive nuclear indices from the photomicrographs.

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Authors and Affiliations

  1. Department of Biological Sciences and Purdue Cancer Center, Purdue University, West Lafayette, IN, USA

    P P Joshi, M V Kulkarni, B K Yu, K R Smith, D L Norton & D S Franklin

  2. Department of Metabolic and Endocrine Diseases, University Medical Center Utrecht, Utrecht, The Netherlands

    W van Veelen & J W M Höppener

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Correspondence to D S Franklin.

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Joshi, P., Kulkarni, M., Yu, B. et al. Simultaneous downregulation of CDK inhibitors p18Ink4c and p27Kip1 is required for MEN2A-RET-mediated mitogenesis. Oncogene 26, 554–570 (2007). https://doi.org/10.1038/sj.onc.1209811

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