Abstract
Malignant melanomas often harbor activating mutations in BRAF (V600E) or, less frequently, in NRAS (Q61R). Intriguingly, the same mutations have been detected at higher incidences in benign nevi, which are largely composed of senescent melanocytes. Overexpression of BRAFV600E or NRASQ61R in human melanocytes in vitro has been shown to induce senescence, although via different mechanisms. How oncogene-induced senescence is overcome during melanoma progression remains unclear. Here, we report that in the majority of analysed BRAFV600E- or NRASQ61R-expressing melanoma cells, C-MYC depletion induced different yet overlapping sets of senescence phenotypes that are characteristic of normal melanocytes undergoing senescence due to overexpression of BRAFV600E or NRASQ61R, respectively. These senescence phenotypes were p16INK4A- or p53-independent, however, several of them were suppressed by genetic or pharmacological inhibition of BRAFV600E or phosphoinositide 3-kinase pathways, including rapamycin-mediated inhibition of mTOR-raptor in NRASQ61R-expressing melanoma cells. Reciprocally, overexpression of C-MYC in normal melanocytes suppressed BRAFV600E-induced senescence more efficiently than NRASQ61R-induced senescence, which agrees with the generally higher rates of activating mutations in BRAF than NRAS gene in human cutaneous melanomas. Our data suggest that one of the major functions of C-MYC overexpression in melanoma progression is to continuous suppress BRAFV600E- or NRASQ61R-dependent senescence programs.
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References
Arvanitis C, Felsher DW . (2006). Conditional transgenic models define how MYC initiates and maintains tumorigenesis. Semin Cancer Biol 16: 313–317.
Bauer J, Curtin JA, Pinkel D, Bastian BC . (2007). Congenital melanocytic nevi frequently harbor NRAS mutations but no BRAF mutations. J Invest Dermatol 127: 179–182.
Benanti JA, Galloway DA . (2004). Normal human fibroblasts are resistant to RAS-induced senescence. Mol Cell Biol 24: 2842–2852.
Biroccio A, Amodei S, Antonelli A, Benassi B, Zupi G . (2003). Inhibition of c-Myc oncoprotein limits the growth of human melanoma cells by inducing cellular crisis. J Biol Chem 278: 35693–35701.
Boehm JS, Hession MT, Bulmer SE, Hahn WC . (2005). Transformation of human and murine fibroblasts without viral oncoproteins. Mol Cell Biol 25: 6464–6474.
Bringold F, Serrano M . (2000). Tumor suppressors and oncogenes in cellular senescence. Exp Gerontol 35: 317–329.
Chin L, Garraway LA, Fisher DE . (2006). Malignant melanoma: genetics and therapeutics in the genomic era. Genes Dev 20: 2149–2182.
Collado M, Serrano M . (2006). The power and the promise of oncogene-induced senescence markers. Nat Rev Cancer 6: 472–476.
Curtin JA, Fridlyand J, Kageshita T, Patel HN, Busam KJ, Kutzner H et al. (2005). Distinct sets of genetic alterations in melanoma. N Engl J Med 353: 2135–2147.
Denoyelle C, Abou-Rjaily G, Bezrookove V, Verhaegen M, Johnson TM, Fullen DR et al. (2006). Anti-oncogenic role of the endoplasmic reticulum differentially activated by mutations in the MAPK pathway. Nat Cell Biol 8: 1053–1063.
Drayton S, Rowe J, Jones R, Vatcheva R, Cuthbert-Heavens D, Marshall J et al. (2003). Tumor suppressor p16INK4a determines sensitivity of human cells to transformation by cooperating cellular oncogenes. Cancer Cell 4: 301–310.
el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM et al. (1993). WAF1, a potential mediator of p53 tumor suppression. Cell 75: 817–825.
Gil J, Kerai P, Lleonart M, Bernard D, Cigudosa JC, Peters G et al. (2005). Immortalization of primary human prostate epithelial cells by c-Myc. Cancer Res 65: 2179–2185.
Gil J, Peters G . (2006). Regulation of the INK4b-ARF-INK4a tumour suppressor locus: all for one or one for all. Nat Rev Mol Cell Biol 7: 667–677.
Goding CR . (2000). Melanocyte development and malignant melanoma. Forum (Genova) 10: 176–187.
Gollob JA, Wilhelm S, Carter C, Kelley SL . (2006). Role of Raf kinase in cancer: therapeutic potential of targeting the Raf/MEK/ERK signal transduction pathway. Semin Oncol 33: 392–406.
Grandori C, Wu KJ, Fernandez P, Ngouenet C, Grim J, Clurman BE et al. (2003). Werner syndrome protein limits MYC-induced cellular senescence. Genes Dev 17: 1569–1574.
Gray-Schopfer V, Wellbrock C, Marais R . (2007). Melanoma biology and new targeted therapy. Nature 445: 851–857.
Greulich KM, Utikal J, Peter RU, Krähn G . (2000). c-MYC and nodular malignant melanoma. A case report. Cancer 89: 97–103.
Guney I, Wu S, Sedivy JM . (2006). Reduced c-Myc signaling triggers telomere-independent senescence by regulating Bmi-1 and p16(INK4a). Proc Natl Acad Sci USA 103: 3645–3650.
Ha L, Ichikawa T, Anver M, Dickins R, Lowe S, Sharpless NE et al. (2007). ARF functions as a melanoma tumor suppressor by inducing p53-independent senescence. Proc Natl Acad Sci USA 104: 10968–10973.
Haluska F, Pemberton T, Ibrahim N, Kalinsky K . (2007). The RTK/RAS/BRAF/PI3K pathways in melanoma: biology, small molecule inhibitors, and potential applications. Semin Oncol 34: 546–554.
Haluska FG, Tsao H, Wu H, Haluska FS, Lazar A, Goel V . (2006). Genetic alterations in signaling pathways in melanoma. Clin Cancer Res 12: 2301s–2307s.
Itahana K, Campisi J, Dimri GP . (2007). Methods to detect biomarkers of cellular senescence: the senescence-associated beta-galactosidase assay. Methods Mol Biol 371: 21–31.
Kastan MB . (2007). Wild-type p53: tumors can't stand it. Cell 128: 837–840.
Kim R, Emi M, Tanabe K, Murakami S . (2006). Role of the unfolded protein response in cell death. Apoptosis 11: 5–13.
Kraehn GM, Utikal J, Udart M, Greulich KM, Bezold G, Kaskel P et al. (2001). Extra c-myc oncogene copies in high risk cutaneous malignant melanoma and melanoma metastases. Br J Cancer 84: 72–79.
Lowe SW, Ruley HE, Jacks T, Housman DE . (1993). p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell 74: 957–967.
Lutz W, Leon J, Eilers M . (2002). Contributions of Myc to tumorigenesis. Biochim Biophys Acta 1602: 61–71.
Maldonado JL, Fridlyand J, Patel H, Jain AN, Busam K, Kageshita T et al. (2003). Determinants of BRAF mutations in primary melanomas. J Natl Cancer Inst 95: 1878–1890.
Meier F, Schittek B, Busch S, Garbe C, Smalley K, Satyamoorthy K et al. (2005). The RAS/RAF/MEK/ERK and PI3K/AKT signaling pathways present molecular targets for the effective treatment of advanced melanoma. Front Biosci 10: 2986–3001.
Michaloglou C, Vredeveld LC, Soengas MS, Denoyelle C, Kuilman T, van der Horst CM et al. (2005). BRAFE600-associated senescence-like cell cycle arrest of human naevi. Nature 436: 720–724.
Mooi WJ, Peeper DS . (2006). Oncogene-induced cell senescence—halting on the road to cancer. N Engl J Med 355: 1037–1046.
Narita M, Nũnez S, Heard E, Narita M, Lin AW, Hearn SA et al. (2003). Rb-mediated heterochromatin formation and silencing of E2F target genes during cellular senescence. Cell 113: 703–716.
Nesbit CE, Tersak JM, Prochownik EV . (1999). MYC oncogenes and human neoplastic disease. Oncogene 18: 3004–3016.
Nikiforov MA, Chandriani S, Park J, Kotenko I, Matheos D, Johnsson A et al. (2002). TRRAP-dependent and TRRAP-independent transcriptional activation by Myc family oncoproteins. Mol Cell Biol 22: 5054–5063.
Pollock PM, Harper UL, Hansen KS, Yudt LM, Stark M, Robbins CM et al. (2003). High frequency of BRAF mutations in nevi. Nat Genet 33: 19–20.
Ross DA, Wilson GD . (1998). Expression of c-myc oncoprotein represents a new prognostic marker in cutaneous melanoma. Br J Surg 85: 46–51.
Sharma A, Trivedi NR, Zimmerman MA, Tuveson DA, Smith CD, Robertson GP . (2005). Mutant V599EB-Raf regulates growth and vascular development of malignant melanoma tumors. Cancer Res 65: 2412–2421.
Sharpless E, Chin L . (2003). The INK4a/ARF locus and melanoma. Oncogene 22: 3092–3098.
Soengas MS, Capodieci P, Polsky D, Mora J, Esteller M, Opitz-Araya X et al. (2001). Inactivation of the apoptosis effector Apaf-1 in malignant melanoma. Nature 409: 207–211.
Takaoka M, Harada H, Deramaudt TB, Oyama K, Andl CD, Johnstone CN et al. (2004). Ha-Ras(G12V) induces senescence in primary and immortalized human esophageal keratinocytes with p53 dysfunction. Oncogene 23: 6760–6768.
Tanaka H, Arakawa H, Yamaguchi T, Shiraishi K, Fukuda S, Matsui K et al. (2004). A ribonucleotide reductase gene involved in a p53-dependent cell-cycle checkpoint for DNA damage. Nature 404: 42–49.
Ventura A, Kirsch DG, McLaughlin ME, Tuveson DA, Grimm J, Lintault L et al. (2007). Restoration of p53 function leads to tumour regression in vivo. Nature 445: 661–665.
Verhaegen M, Bauer JA, Martin de la Vega C, Wang G, Wolter KG, Brenner JC et al. (2006). A novel BH3 mimetic reveals a mitogen-activated protein kinase-dependent mechanism of melanoma cell death controlled by p53 and reactive oxygen species. Cancer Res 66: 11348–11359.
Vita M, Henriksson M . (2006). The Myc oncoprotein as a therapeutic target for human cancer. Semin Cancer Biol 16: 318–330.
Wang H, Mannava S, Grachtchouk V, Zhuang D, Soengas MS, Gudkov AV et al. (2008). c-Myc depletion inhibits proliferation of human tumor cells at various stages of the cell cycle. Oncogene 27: 1905–1915.
Wu CH, van Riggelen J, Yetil A, Fan AC, Bachireddy P, Felsher DW . (2007). Cellular senescence is an important mechanism of tumor regression upon c-Myc inactivation. Proc Natl Acad Sci USA 104: 13028–13033.
Xue W, Zender L, Miething C, Dickins RA, Hernando E, Krizhanovsky V et al. (2007). Senescence and tumour clearance is triggered by p53 restoration in murine liver carcinomas. Nature 445: 656–660.
Yang Q, Guan KL . (2007). Expanding mTOR signaling. Cell Res 17: 666–681.
Acknowledgements
We are grateful to Michelle Vinco for assistance with the human tissue work. This work was supported by Grants NIH R01 CA120244 (MAN), CA107237 (MSS), and, in part, by the University of Michigan's Cancer Center Support Grant (NIH 5 P30 CA46592). MAN is a Melanoma Research Foundation Scholar.
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Zhuang, D., Mannava, S., Grachtchouk, V. et al. C-MYC overexpression is required for continuous suppression of oncogene-induced senescence in melanoma cells. Oncogene 27, 6623–6634 (2008). https://doi.org/10.1038/onc.2008.258
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DOI: https://doi.org/10.1038/onc.2008.258
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