Loss of p16Ink4a confers susceptibility to metastatic melanoma in mice

Abstract

CDKN2A (INK4a/ARF) is frequently disrupted in various types of human cancer, and germline mutations of this locus can confer susceptibility to melanoma and other tumours1. However, because CDKN2A encodes two distinct cell cycle inhibitory proteins, p16INK4a and p14ARF (p19Arf in mice)2, the mechanism of tumour suppression by CDKN2A has remained controversial. Genetic disruption of Cdkn2a(p19Arf) (hereafter Arf) alone predisposes mice to tumorigenesis3, demonstrating that Arf is a tumour-suppressor gene in mice. We mutated mice specifically in Cdkn2a(p16Ink4a) (hereafter Ink4a). Here we demonstrate that these mice, designated Ink4a*/*, do not show a significant predisposition to spontaneous tumour formation within 17 months. Embryo fibroblasts derived from them proliferate normally, are mortal, and are not transformed by oncogenic HRAS. The very mild phenotype of the Ink4a*/* mice implies that the very strong phenotypes of the original Ink4a/ArfΔ2,3 mice were primarily or solely due to loss of Arf. However, Ink4a*/Δ2,3 mice that are deficient for Ink4a and heterozygous for Arf spontaneously develop a wide spectrum of tumours, including melanoma. Treatment of these mice with the carcinogen 7,12-dimethylbenzanthracene (DMBA) results in an increased incidence of melanoma, with frequent metastases. Our results show that, in the mouse, Ink4a is a tumour-suppressor gene that, when lost, can recapitulate the tumour predisposition seen in humans.

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Figure 1: Generation of Ink4a mutant mice.
Figure 2: Growth characteristics of Ink4a*/* MEFs.
Figure 3: Ink4a is a tumour-suppressor gene.
Figure 4: Histology of melanocytic tumours in Ink4a*/Δ2,3 mice.

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Acknowledgements

We thank D. Peeper and M. van Lohuizen for advice and retroviral vectors, M. van der Valk for histological analysis, K. Rajewsky for the Deleter Cre mice, C. Sherr for Arf-/- MEFs, G. Nolan for ΦNX-E retroviral producer cells, and J. Vink and K. van Veen for technical assistance, and the Netherlands Cancer Institute animal facility for providing animal care. Part of this work was supported by the Dutch Cancer Society.

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Correspondence to Anton Berns.

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