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Adhesion control of cyclin D1 and p27Kip1 levels is deregulated in melanoma cells through BRAF-MEK-ERK signaling

Oncogene volume 24, pages 3459–3471 (2005)Cite this article

Abstract

Mutations in BRAF, a component of extracellular signal-regulated kinases 1 and 2 (ERK) cascade, are frequent in melanoma. It is important to understand how BRAF mutations contribute to malignant traits including anchorage- and growth factor-independence. We have previously shown that efficient activation of ERK in normal human epidermal melanocytes (NHEM) requires both adhesion to the extracellular matrix and growth factors. Mutant V599E BRAF is sufficient to promote ERK activation independent of adhesion and growth factors. Here, we analysed regulation of G1 cell cycle events in NHEM and human melanoma cells. We show that S phase entry in NHEM requires both adhesion and growth factor signaling through the MEK-ERK pathway. This control correlates with induction of cyclin D1 and downregulation of p27Kip1, two key G1 cell cycle events. In melanoma cells expressing V599E BRAF, cyclin D1 was constitutively expressed independent of adhesion but dependent upon MEK activation and nuclear accumulation of ERK. Reduction of cyclin D1 levels by RNA interference inhibited S phase entry in melanoma cells. Importantly, expression of V599E BRAF in NHEM was sufficient to promote cyclin D1 promoter activity in the absence of adhesion. Additionally, p27Kip1 levels were downregulated in V599E BRAF-expressing melanoma cells and active BRAF was sufficient to downregulate p27Kip1 in serum-starved NHEM. Thus, adhesion-growth factor cooperation, leading to efficient activation of ERK, regulates cyclin D1 and p27Kip1 levels in human melanocytes and mutant BRAF overrides adhesion-growth factor control of these two G1 cell cycle proteins in melanomas. These findings provide important insight into how BRAF mutations contribute to aberrant human melanocyte proliferation.

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Abbreviations

CDK:

cyclin-dependent kinase

ECM:

extracellular matrix

ERK:

extracellular signal-regulated kinases 1 and 2

MAPK:

mitogen-activated protein kinase

MEK:

MAPK/ERK kinase

NHEM:

normal human epidermal melanocytes

RGP:

radial growth phase

TPA:

12-O-tetradecanoyl phorbol 13-acetate

VGP:

vertical growth phase

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Acknowledgements

We are indebted to the staff of The Birth Place at Albany Medical Center for providing discarded tissue samples for melanocyte isolation. We are very grateful to Drs Alan Howe, Stephen Keyse, Richard Marais and Martin McMahon for cDNA constructs and Dr Meenhard Herlyn for melanoma cell lines. Also, we thank Dr Michael DiPersio (Center for Cell Biology and Cancer Research, Albany Medical College) for his critical reading of the manuscript. This work was supported by National Institutes of Health Grants GM067893 (to AEA), CA81419 (to KP), and an institutional predoctoral training grant (T32-HL-07194).

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  1. Center for Cell Biology and Cancer Research, Albany Medical College, 47 New Scotland Avenue, MC-165, Albany, NY, 12208, USA

    Kavita V Bhatt, Laurie S Spofford, Gazelle Aram, Meghan McMullen, Kevin Pumiglia & Andrew E Aplin

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  1. Kavita V Bhatt
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Correspondence to Andrew E Aplin.

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Bhatt, K., Spofford, L., Aram, G. et al. Adhesion control of cyclin D1 and p27Kip1 levels is deregulated in melanoma cells through BRAF-MEK-ERK signaling. Oncogene 24, 3459–3471 (2005). https://doi.org/10.1038/sj.onc.1208544

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