Abstract
KIT mutations are frequent in acral, mucosal and chronic sun-damage (CSD) melanoma, but little is known about the mechanisms driving the transformation of KIT-mutated melanocytes into melanoma cells. We showed that exposition of melanocytes harboring the L576PKIT mutation to a hypoxic environment induced their transformation into malignant cells. Transformed L576PKIT melanocytes showed downregulation of MITF expression characteristic of melanoma initiating cells (MICs). In agreement, these cells were able to form spheres in neural crest cell medium and low-adherence conditions, also a characteristic of MICs. Downregulation of MITF by RNA interference induced transformation of KIT-mutated melanocytes in normoxia and acquisition of a MIC phenotype by these cells. Hence, low level of MITF cooperates with oncogenic KIT to transform melanocytes. Activation of the cAMP pathway in transformed L576PKIT melanocytes stimulated MITF expression, and reduced cellular proliferation and sphere formation. These findings highlight the essential role of MITF in revealing the oncogenic activity of KIT in melanocytes and suggest that the cAMP pathway is a therapeutic target in KIT-mutated melanoma.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Liu H, Chen X, Focia PJ, He X . Structural basis for stem cell factor-KIT signaling and activation of class III receptor tyrosine kinases. EMBO J 2007; 26: 891–901.
Ronnstrand L . Signal transduction via the stem cell factor receptor/c-Kit. Cell Mol Life Sci 2004; 61: 2535–2548.
Young SM, Cambareri AC, Odell A, Geary SM, Ashman LK . Early myeloid cells expressing c-KIT isoforms differ in signal transduction, survival and chemotactic responses to Stem Cell Factor. Cell Signal 2007; 19: 2572–2581.
Garrido MC, Bastian BC . KIT as a therapeutic target in melanoma. J Invest Dermatol 2010; 130: 20–27.
Giebel LB, Spritz RA . Mutation of the KIT (mast/stem cell growth factor receptor) protooncogene in human piebaldism. Proc Natl Acad Sci USA 1991; 88: 8696–8699.
Huang S, Luca M, Gutman M, McConkey DJ, Langley KE, Lyman SD et al. Enforced c-KIT expression renders highly metastatic human melanoma cells susceptible to stem cell factor-induced apoptosis and inhibits their tumorigenic and metastatic potential. Oncogene 1996; 13: 2339–2347.
Lassam N, Bickford S . Loss of c-kit expression in cultured melanoma cells. Oncogene 1992; 7: 51–56.
Curtin JA, Busam K, Pinkel D, Bastian BC . Somatic activation of KIT in distinct subtypes of melanoma. J Clin Oncol 2006; 24: 4340–4346.
Curtin JA, Fridlyand J, Kageshita T, Patel HN, Busam KJ, Kutzner H et al. Distinct sets of genetic alterations in melanoma. N Engl J Med 2005; 353: 2135–2147.
Bastian BC, Esteve-Puig R . Targeting activated KIT signaling for melanoma therapy. J Clin Oncol 2013; 31: 3288–3290.
Park E, Yang S, Emley A, DeCarlo K, Richards J, Mahalingam M . Lack of correlation between immunohistochemical expression of CKIT and KIT mutations in atypical acral nevi. Am J Dermatopathol 2012; 34: 41–46.
Monsel G, Ortonne N, Bagot M, Bensussan A, Dumaz N . c-Kit mutants require hypoxia-inducible factor 1alpha to transform melanocytes. Oncogene 2010; 29: 227–236.
Evans SM, Schrlau AE, Chalian AA, Zhang P, Koch CJ . Oxygen levels in normal and previously irradiated human skin as assessed by EF5 binding. J Invest Dermatol 2006; 126: 2596–2606.
Bedogni B, Welford SM, Cassarino DS, Nickoloff BJ, Giaccia AJ, Powell MB . The hypoxic microenvironment of the skin contributes to Akt-mediated melanocyte transformation. Cancer Cell 2005; 8: 443–454.
Horikoshi T, Balin AK, Carter DM . Effects of oxygen tension on the growth and pigmentation of normal human melanocytes. J Invest Dermatol 1991; 96: 841–844.
Rezvani HR, Ali N, Nissen LJ, Harfouche G, de Verneuil H, Taieb A et al. HIF-1alpha in epidermis: oxygen sensing, cutaneous angiogenesis, cancer, and non-cancer disorders. J Invest Dermatol 2011; 131: 1793–1805.
Bedogni B, Powell MB . Hypoxia, melanocytes and melanoma - survival and tumor development in the permissive microenvironment of the skin. Pigment Cell Melanoma Res 2009; 22: 166–174.
Mohyeldin A, Garzon-Muvdi T, Quinones-Hinojosa A . Oxygen in stem cell biology: a critical component of the stem cell niche. Cell Stem Cell 2010; 7: 150–161.
Cheli Y, Giuliano S, Botton T, Rocchi S, Hofman V, Hofman P et al. Mitf is the key molecular switch between mouse or human melanoma initiating cells and their differentiated progeny. Oncogene 2011; 30: 2307–2318.
Fang D, Nguyen TK, Leishear K, Finko R, Kulp AN, Hotz S et al. A tumorigenic subpopulation with stem cell properties in melanomas. Cancer Res 2005; 65: 9328–9337.
Kumar SM, Liu S, Lu H, Zhang H, Zhang PJ, Gimotty PA et al. Acquired cancer stem cell phenotypes through Oct4-mediated dedifferentiation. Oncogene 2012; 31: 4898–4911.
Ramgolam K, Lauriol J, Lalou C, Lauden L, Michel L, de la Grange P et al. Melanoma spheroids grown under neural crest cell conditions are highly plastic migratory/invasive tumor cells endowed with immunomodulator function. PLoS One 2011; 6: e18784.
Schatton T, Murphy GF, Frank NY, Yamaura K, Waaga-Gasser AM, Gasser M et al. Identification of cells initiating human melanomas. Nature 2008; 451: 345–349.
Cheli Y, Giuliano S, Fenouille N, Allegra M, Hofman V, Hofman P et al. Hypoxia and MITF control metastatic behaviour in mouse and human melanoma cells. Oncogene 2012; 31: 2461–2470.
Feige E, Yokoyama S, Levy C, Khaled M, Igras V, Lin RJ et al. Hypoxia-induced transcriptional repression of the melanoma-associated oncogene MITF. Proc Natl Acad Sci USA 2011; 108: E924–E933.
Hoek KS, Goding CR . Cancer stem cells versus phenotype-switching in melanoma. Pigment Cell Melanoma Res 2010; 23: 746–759.
Dumaz N, Andre J, Sadoux A, Laugier F, Podgorniak MP, Mourah S et al. Driver KIT mutations in melanoma cluster in four hotspots. Melanoma research 2015; 25: 88–90.
Bourillon A, Hu HH, Hetet G, Lacapere JJ, Andre J, Descamps V et al. Genetic variation at KIT locus may predispose to melanoma. Pigment Cell Melanoma Res 2013; 26: 88–96.
Liang R, Wallace AR, Schadendorf D, Rubin BP . The phosphatidyl inositol 3-kinase pathway is central to the pathogenesis of Kit-activated melanoma. Pigment Cell Melanoma Res 2011; 24: 714–723.
Todd JR, Scurr LL, Becker TM, Kefford RF, Rizos H . The MAPK pathway functions as a redundant survival signal that reinforces the PI3K cascade in c-Kit mutant melanoma. Oncogene 2012; 33: 236–245.
Ohanna M, Cheli Y, Bonet C, Bonazzi VF, Allegra M, Giuliano S et al. Secretome from senescent melanoma engages the STAT3 pathway to favor reprogramming of naive melanoma towards a tumor-initiating cell phenotype. Oncotarget 2013; 4: 2212–2224.
Hamai A, Richon C, Meslin F, Faure F, Kauffmann A, Lecluse Y et al. Imatinib enhances human melanoma cell susceptibility to TRAIL-induced cell death: Relationship to Bcl-2 family and caspase activation. Oncogene 2006; 25: 7618–7634.
Garraway LA, Widlund HR, Rubin MA, Getz G, Berger AJ, Ramaswamy S et al. Integrative genomic analyses identify MITF as a lineage survival oncogene amplified in malignant melanoma. Nature 2005; 436: 117–122.
von Euw E, Atefi M, Attar N, Chu C, Zachariah S, Burgess BL et al. Antitumor effects of the investigational selective MEK inhibitor TAK733 against cutaneous and uveal melanoma cell lines. Mol Cancer 2012; 11: 22.
Carvajal RD, Antonescu CR, Wolchok JD, Chapman PB, Roman RA, Teitcher J et al. KIT as a therapeutic target in metastatic melanoma. JAMA 2011; 305: 2327–2334.
Guo J, Si L, Kong Y, Flaherty KT, Xu X, Zhu Y et al. Phase II, open-label, single-arm trial of imatinib mesylate in patients with metastatic melanoma harboring c-Kit mutation or amplification. J Clin Oncol 2011; 29: 2904–2909.
Hodi FS, Corless CL, Giobbie-Hurder A, Fletcher JA, Zhu M, Marino-Enriquez A et al. Imatinib for Melanomas Harboring Mutationally Activated or Amplified KIT Arising on Mucosal, Acral, and Chronically Sun-Damaged Skin. J Clin Oncol 2013; 31: 3182–3190.
Marquette A, Andre J, Bagot M, Bensussan A, Dumaz N ERK . and PDE4 cooperate to induce RAF isoform switching in melanoma. Nat Struct Mol Biol 2011; 18: 584–591.
Bennett DC, Cooper PJ, Hart IR . A line of non-tumorigenic mouse melanocytes, syngeneic with the B16 melanoma and requiring a tumour promoter for growth. Int J Cancer 1987; 39: 414–418.
Acknowledgements
We thank Pr Antoni Ribas for the M230 melanoma cell line. This work was funded by L’Oréal Research & Innovation, INSERM, Université Paris Diderot and Fondation ARC pour la Recherche sur le Cancer. FL was supported by L’Oréal Research & Innovation.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies this paper on the Oncogene website
Supplementary information
Rights and permissions
About this article
Cite this article
Laugier, F., Delyon, J., André, J. et al. Hypoxia and MITF regulate KIT oncogenic properties in melanocytes. Oncogene 35, 5070–5077 (2016). https://doi.org/10.1038/onc.2016.39
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2016.39