Abstract
Most normal mammalian cells have a finite lifespan1, thought to constitute a protective mechanism against unlimited proliferation2,3,4. This phenomenon, called senescence, is driven by telomere attrition, which triggers the induction of tumour suppressors including p16INK4a (ref. 5). In cultured cells, senescence can be elicited prematurely by oncogenes6; however, whether such oncogene-induced senescence represents a physiological process has long been debated. Human naevi (moles) are benign tumours of melanocytes that frequently harbour oncogenic mutations (predominantly V600E, where valine is substituted for glutamic acid) in BRAF7, a protein kinase and downstream effector of Ras. Nonetheless, naevi typically remain in a growth-arrested state for decades and only rarely progress into malignancy (melanoma)8,9,10. This raises the question of whether naevi undergo BRAFV600E-induced senescence. Here we show that sustained BRAFV600E expression in human melanocytes induces cell cycle arrest, which is accompanied by the induction of both p16INK4a and senescence-associated acidic β-galactosidase (SA-β-Gal) activity, a commonly used senescence marker. Validating these results in vivo, congenital naevi are invariably positive for SA-β-Gal, demonstrating the presence of this classical senescence-associated marker in a largely growth-arrested, neoplastic human lesion. In growth-arrested melanocytes, both in vitro and in situ, we observed a marked mosaic induction of p16INK4a, suggesting that factors other than p16INK4a contribute to protection against BRAFV600E-driven proliferation. Naevi do not appear to suffer from telomere attrition, arguing in favour of an active oncogene-driven senescence process, rather than a loss of replicative potential. Thus, both in vitro and in vivo, BRAFV600E-expressing melanocytes display classical hallmarks of senescence, suggesting that oncogene-induced senescence represents a genuine protective physiological process.
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References
Hayflick, L. The limited in vitro lifetime of human diploid cell strains. Exp. Cell Res. 37, 614–636 (1965)
Mathon, N. F. & Lloyd, A. C. Cell senescence and cancer. Nature Rev. Cancer 1, 203–213 (2001)
Lowe, S. W., Cepero, E. & Evan, G. Intrinsic tumour suppression. Nature 432, 307–315 (2004)
Campisi, J. Senescent cells, tumour suppression, and organismal aging: good citizens, bad neighbors. Cell 120, 513–522 (2005)
Shay, J. W. & Roninson, I. B. Hallmarks of senescence in carcinogenesis and cancer therapy. Oncogene 23, 2919–2933 (2004)
Serrano, M., Lin, A. W., McCurrach, M. E., Beach, D. & Lowe, S. W. Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88, 593–602 (1997)
Pollock, P. M. et al. High frequency of BRAF mutations in nevi. Nature Genet. 33, 19–20 (2003)
Kuwata, T., Kitagawa, M. & Kasuga, T. Proliferative activity of primary cutaneous melanocytic tumours. Virchows Arch. A Pathol. Anat. Histopathol. 423, 359–364 (1993)
Bennett, D. C. Human melanocyte senescence and melanoma susceptibility genes. Oncogene 22, 3063–3069 (2003)
Chin, L., Merlino, G. & DePinho, R. A. Malignant melanoma: modern black plague and genetic black box. Genes Dev. 12, 3467–3481 (1998)
Robinson, W. A. et al. Human acquired naevi are clonal. Melanoma Res. 8, 499–503 (1998)
Davies, H. et al. Mutations of the BRAF gene in human cancer. Nature 417, 949–954
Wellbrock, C. et al. V599EB-RAF is an oncogene in melanocytes. Cancer Res. 64, 2338–2342 (2004)
Mooi, W. J. & Krausz, T. Biopsy Pathology of Melanocytic Disorders 56–105 (Chapman & Hall Medical, London, 1992)
Dimri, G. P. et al. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc. Natl Acad. Sci. USA 92, 9363–9367 (1995)
Sharpless, E. & Chin, L. The INK4a/ARF locus and melanoma. Oncogene 22, 3092–3098 (2003)
Zhu, J., Woods, D., McMahon, M. & Bishop, J. M. Senescence of human fibroblasts induced by oncogenic Raf. Genes Dev. 12, 2997–3007 (1998)
Lin, A. W. et al. Premature senescence involving p53 and p16 is activated in response to constitutive MEK/MAPK mitogenic signalling. Genes Dev. 12, 3008–3019 (1998)
Narita, M. et al. Rb-mediated heterochromatin formation and silencing of E2F target genes during cellular senescence. Cell 113, 703–716 (2003)
Gruis, N. A. et al. Homozygotes for CDKN2 (p16) germline mutation in Dutch familial melanoma kindreds. Nature Genet. 10, 351–353 (1995)
Bandyopadhyay, D. et al. The human melanocyte: a model system to study the complexity of cellular aging and transformation in non-fibroblastic cells. Exp. Gerontol. 36, 1265–1275 (2001)
Wang, Y. L., Uhara, H., Yamazaki, Y., Nikaido, T. & Saida, T. Immunohistochemical detection of CDK4 and p16INK4 proteins in cutaneous malignant melanoma. Br. J. Dermatol. 134, 269–275 (1996)
Kamb, A. et al. A cell cycle regulator potentially involved in genesis of many tumour types. Science 264, 436–440 (1994)
Beausejour, C. M. et al. Reversal of human cellular senescence: roles of the p53 and p16 pathways. EMBO J. 22, 4212–4222 (2003)
Bastian, B. C. Understanding the progression of melanocytic neoplasia using genomic analysis: from fields to cancer. Oncogene 22, 3081–3086 (2003)
Miracco, C. et al. Quantitative in situ evaluation of telomeres in fluorescence in situ hybridization-processed sections of cutaneous melanocytic lesions and correlation with telomerase activity. Br. J. Dermatol. 146, 399–408 (2002)
Peeper, D. S. & Mooi, W. J. Pathogenesis of melanocytic naevi: growth arrest linked with cellular senescence? Histopathology 41, S139–S143 (2002)
Patton, E. E. et al. BRAF mutations are sufficient to promote nevi formation and cooperate with p53 in the genesis of melanoma. Curr. Biol. 15, 249–254 (2005)
Hingorani, S. R., Jacobetz, M. A., Robertson, G. P., Herlyn, M. & Tuveson, D. A. Suppression of BRAF(V599E) in human melanoma abrogates transformation. Cancer Res. 63, 5198–5202 (2003)
Peeper, D. S. et al. A functional screen identifies hDRIL1 as an oncogene that rescues RAS-induced senescence. Nature Cell Biol. 4, 148–153 (2002)
Acknowledgements
We thank D. Atsma, E. Mesman and J. Zevenhoven for help with immunohistochemistry; S. Douma for analytical support; L. Oomen, L. Brocks and J. van Rheenen for help with microscopy; N. Gruis and C. Out for p16INK4a-deficient fibroblasts; L. Zaal and A. van der Wal for help with obtaining congenital naevus specimens; M. Voorhoeve and R. Agami for pRetroSuper, pRetroSuper-Blasticidin and pRetroSuper-GFP; S. Gryaznov for the telomere probe; R. Beijersbergen and M. van Lohuizen for reagents; G. Abou-Rjaily for help with lentiviral infections; P. Krimpenfort and colleagues in the Peeper laboratory for discussions; R. Bernards for support; and M. van Lohuizen and A. Berns for suggestions and reading of the manuscript. M.S.S is supported by an NIH grant. M.S.S. is a V Foundation for Cancer Research Scholar. L.C.W.V., T.K. and D.S.P. were supported by the Netherlands Organization for Scientific Research (NWO).
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Supplementary information
Supplmentary Figure S1
Ectopic overexpression of BRAFE600 causes a senescence-like cell cycle arrest. (PDF 236 kb)
Supplmentary Figure S2a-f
Physiologic levels of BRAFE600 induce senescence in a p16INK4a-independent manner. (PDF 818 kb)
Supplementary Figure 2g-l
Physiologic levels of BRAFE600 induce senescence in a p16INK4a-independent manner. (PDF 436 kb)
Supplmentary Figure S3
BRAFE600 fails to induce p14ARF. (PDF 23 kb)
Supplementary Figure S4
Sustained knockdown of p16INK4a cause an increase in cellular proliferation rate (PDF 48 kb)
Supplementary Figure S5
BRAFE600 mutational analysis in human congenital nevi. (PDF 39 kb)
Supplementary Figure S6
Mosaic pattern of p16INK4a-positivity in nevi. (PDF 152 kb)
Supplementary Figure S7
No apparent telomere loss in nevi. (PDF 648 kb)
Supplementary Figure Legends
Text to accompany the above Supplementary Figures. (RTF 14 kb)
Supplementary Methods
Additional descriptions of methods used in this study, to accompany those detailed in the main text. (RTF 9 kb)
Supplmentary Table
Tumorigenicity assay. (PDF 42 kb)
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Michaloglou, C., Vredeveld, L., Soengas, M. et al. BRAFE600-associated senescence-like cell cycle arrest of human naevi. Nature 436, 720–724 (2005). https://doi.org/10.1038/nature03890
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DOI: https://doi.org/10.1038/nature03890
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