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BMC cancer2014; 14; 857; doi: 10.1186/1471-2407-14-857

Constitutive activation of the ERK pathway in melanoma and skin melanocytes in Grey horses.

Abstract: Constitutive activation of the ERK pathway, occurring in the vast majority of melanocytic neoplasms, has a pivotal role in melanoma development. Different mechanisms underlie this activation in different tumour settings. The Grey phenotype in horses, caused by a 4.6 kb duplication in intron 6 of Syntaxin 17 (STX17), is associated with a very high incidence of cutaneous melanoma, but the molecular mechanism behind the melanomagenesis remains unknown. Here, we investigated the involvement of the ERK pathway in melanoma development in Grey horses. Methods: Grey horse melanoma tumours, cell lines and normal skin melanocytes were analyzed with help of indirect immunofluorescence and immunoblotting for the expression of phospho-ERK1/2 in comparison to that in non-grey horse and human counterparts. The mutational status of BRAF, RAS, GNAQ, GNA11 and KIT genes in Grey horse melanomas was determined by direct sequencing. The effect of RAS, RAF and PI3K/AKT pathways on the activation of the ERK signaling in Grey horse melanoma cells was investigated with help of specific inhibitors and immunoblotting. Individual roles of RAF and RAS kinases on the ERK activation were examined using si-RNA based approach and immunoblotting. Results: We found that the ERK pathway is constitutively activated in Grey horse melanoma tumours and cell lines in the absence of somatic activating mutations in BRAF, RAS, GNAQ, GNA11 and KIT genes or alterations in the expression of the main components of the pathway. The pathway is mitogenic and is mediated by BRAF, CRAF and KRAS kinases. Importantly, we found high activation of the ERK pathway also in epidermal melanocytes, suggesting a general predisposition to melanomagenesis in these horses. Conclusions: These findings demonstrate that the presence of the intronic 4.6 kb duplication in STX17 is strongly associated with constitutive activation of the ERK pathway in melanocytic cells in Grey horses in the absence of somatic mutations commonly linked to the activation of this pathway during melanomagenesis. These findings are consistent with the universal importance of the ERK pathway in melanomagenesis and may have valuable implications for human melanoma research.
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Publication Date: 2014-11-21 PubMed ID: 25413220PubMed Central: PMC4254013DOI: 10.1186/1471-2407-14-857Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research explores why Grey horses, who have a specific genetic mutation in Syntaxin 17 (STX17), have a high incidence of skin melanoma. The study finds that the ERK pathway, commonly associated with melanoma development, is constitutively activated in these horses. Notably, this happens in the absence of other common melanoma-associated genetic mutations, indicating a universal role of the ERK pathway in melanoma development.

Research Methodology

  • This research primarily involved detailed investigations of melanoma tumors, cell lines, and normal skin melanocytes of Grey horses.
  • Techniques such as indirect immunofluorescence and immunoblotting were used to analyze the expression of phospho-ERK1/2 in comparison with non-grey horses and human samples.
  • Mutation status of genes like BRAF, RAS, GNAQ, GNA11 and KIT associated with melanomas were checked through direct sequencing.
  • Further, the impact of RAS, RAF, and PI3K/AKT pathways on the activation of the ERK signaling in Grey horse melanoma cells was investigated using specific inhibitors and immunoblotting.
  • Lastly, individual roles of RAF and RAS kinases on the ERK activation were analyzed using si-RNA based approach and immunoblotting.

Research Findings

  • The ERK pathway was found to be constitutively activated in Grey horse melanoma tumours and cell lines, even without common somatic mutations in BRAF, RAS, GNAQ, GNA11, and KIT genes or alteration of main pathway components.
  • This pathway was found to be mitogenic and is mediated by BRAF, CRAF, and KRAS kinases.
  • Notably, higher activation of the ERK pathway was also found in epidermal melanocytes, indicating a general predisposition to melanoma in these horses.

Conclusions

  • The study concludes that presence of the intronic 4.6 kb duplication in STX17 gene is strongly linked with constitutive activation of the ERK pathway in Grey horses, even in absence of typically observed somatic mutations linked to melanoma.
  • This suggests the universal importance of the ERK pathway in melanoma development and brings valuable insights for human melanoma research.

Cite This Article

APA
Jiang L, Campagne C, Sundström E, Sousa P, Imran S, Seltenhammer M, Pielberg G, Olsson MJ, Egidy G, Andersson L, Golovko A. (2014). Constitutive activation of the ERK pathway in melanoma and skin melanocytes in Grey horses. BMC Cancer, 14, 857. https://doi.org/10.1186/1471-2407-14-857

Publication

BMC cancer
ISSN: 1471-2407
NlmUniqueID: 100967800
Country: England
Language: English
Volume: 14
Pages: 857

Researcher Affiliations

Jiang, Lin
    Campagne, Cécile
      Sundström, Elisabeth
        Sousa, Pedro
          Imran, Saima
            Seltenhammer, Monika
              Pielberg, Gerli
                Olsson, Mats J
                  Egidy, Giorgia
                    Andersson, Leif
                      Golovko, Anna
                      • Science for Life Laboratory, Department of Medical Biochemistry and Microbiology, Uppsala University, Box 582, SE-751 23 Uppsala, Sweden. Anna.Golovko@imbim.uu.se.

                      MeSH Terms

                      • Animals
                      • Cell Line, Tumor
                      • Genetic Variation
                      • Genotype
                      • Horse Diseases / genetics
                      • Horse Diseases / metabolism
                      • Horses
                      • Humans
                      • MAP Kinase Signaling System
                      • Melanocytes / metabolism
                      • Melanoma / veterinary
                      • Mitogen-Activated Protein Kinase 1 / metabolism
                      • Mitogen-Activated Protein Kinase 3 / metabolism
                      • Oncogene Proteins / genetics
                      • Proto-Oncogene Proteins B-raf / genetics
                      • Proto-Oncogene Proteins p21(ras) / genetics

                      References

                      This article includes 33 references
                      1. Smalley KS. A pivotal role for ERK in the oncogenic behaviour of malignant melanoma?. Int J Cancer 2003;104(5):527–532.
                        doi: 10.1002/ijc.10978pubmed: 12594806google scholar: lookup
                      2. Fecher LA, Amaravadi RK, Flaherty KT. The MAPK pathway in melanoma. Curr Opin Oncol 2008;20(2):183–189.
                        doi: 10.1097/CCO.0b013e3282f5271cpubmed: 18300768google scholar: lookup
                      3. Hearing V, Leong S. From Melanocytes to Melanoma: The Progression to Malignancy. Totowa, NJ: Humana Press; 2006.
                      4. Davies H, Bignell GR, Cox C, Stephens P, Edkins S, Clegg S, Teague J, Woffendin H, Garnett MJ, Bottomley W, Davis N, Dicks E, Ewing R, Floyd Y, Gray K, Hall S, Hawes R, Hughes J, Kosmidou V, Menzies A, Mould C, Parker A, Stevens C, Watt S, Hooper S, Wilson R, Jayatilake H, Gusterson BA, Cooper C, Shipley J. Mutations of the BRAF gene in human cancer. Nature 2002;417(6892):949–954.
                        doi: 10.1038/nature00766pubmed: 12068308google scholar: lookup
                      5. Van Raamsdonk CD, Bezrookove V, Green G, Bauer J, Gaugler L, O’Brien JM, Simpson EM, Barsh GS, Bastian BC. Frequent somatic mutations of GNAQ in uveal melanoma and blue naevi. Nature 2009;457(7229):599–602.
                        doi: 10.1038/nature07586pmc: PMC2696133pubmed: 19078957google scholar: lookup
                      6. Curtin JA, Busam K, Pinkel D, Bastian BC. Somatic activation of KIT in distinct subtypes of melanoma. J Clin Oncol 2006;24(26):4340–4346.
                        doi: 10.1200/JCO.2006.06.2984pubmed: 16908931google scholar: lookup
                      7. Jiang X, Zhou J, Yuen NK, Corless CL, Heinrich MC, Fletcher JA, Demetri GD, Widlund HR, Fisher DE, Hodi FS. Imatinib targeting of KIT-mutant oncoprotein in melanoma. Clin Cancer Res 2008;14(23):7726–7732.
                        doi: 10.1158/1078-0432.CCR-08-1144pubmed: 19047099google scholar: lookup
                      8. Monsel G, Ortonne N, Bagot M, Bensussan A, Dumaz N. c-Kit mutants require hypoxia-inducible factor 1alpha to transform melanocytes. Oncogene 2010;29(2):227–236.
                        doi: 10.1038/onc.2009.320pubmed: 19802003google scholar: lookup
                      9. Nikolaev SI, Rimoldi D, Iseli C, Valsesia A, Robyr D, Gehrig C, Harshman K, Guipponi M, Bukach O, Zoete V, Michielin O, Muehlethaler K, Speiser D, Beckmann JS, Xenarios I, Halazonetis TD, Jongeneel CV, Stevenson BJ, Antonarakis SE. Exome sequencing identifies recurrent somatic MAP2K1 and MAP2K2 mutations in melanoma. Nat Genet 2012;44(2):133–139.
                        doi: 10.1038/ng.1026pubmed: 22197931google scholar: lookup
                      10. Tanami H, Imoto I, Hirasawa A, Yuki Y, Sonoda I, Inoue J, Yasui K, Misawa-Furihata A, Kawakami Y, Inazawa J. Involvement of overexpressed wild-type BRAF in the growth of malignant melanoma cell lines. Oncogene 2004;23(54):8796–8804.
                        doi: 10.1038/sj.onc.1208152pubmed: 15467732google scholar: lookup
                      11. Panka DJ, Atkins MB, Mier JW. Targeting the mitogen-activated protein kinase pathway in the treatment of malignant melanoma. Clin Cancer Res 2006;12(72):2371s–2375s.
                        doi: 10.1158/1078-0432.CCR-05-2539pubmed: 16609061google scholar: lookup
                      12. Tsavachidou D, Coleman ML, Athanasiadis G, Li S, Licht JD, Olson MF, Weber BL. SPRY2 is an inhibitor of the ras/extracellular signal-regulated kinase pathway in melanocytes and melanoma cells with wild-type BRAF but not with the V599E mutant. Cancer Res 2004;64(16):5556–5559.
                        doi: 10.1158/0008-5472.CAN-04-1669pubmed: 15313890google scholar: lookup
                      13. Sutton RH, Coleman GT. Melanoma and the Graying Horse. Barton, Australia: RIRDC Research Paper Series; 1997. pp. 1–34.
                      14. Smith SH, Goldschmidt MH, McManus PM. A comparative review of melanocytic neoplasms. Vet Pathol 2002;39(6):651–678.
                        doi: 10.1354/vp.39-6-651pubmed: 12450197google scholar: lookup
                      15. Gorham S, Robl M. Melanoma in the gray horse: the darker side of equine aging. Vet Med 1986;11:1191–1204.
                      16. Seltenhammer MH, Heere-Ress E, Brandt S, Druml T, Jansen B, Pehamberger H, Niebauer GW. Comparative histopathology of grey-horse-melanoma and human malignant melanoma. Pigment Cell Res 2004;17(6):674–681.
                        doi: 10.1111/j.1600-0749.2004.00192.xpubmed: 15541026google scholar: lookup
                      17. Rosengren Pielberg G, Golovko A, Sundstrom E, Curik I, Lennartsson J, Seltenhammer MH, Druml T, Binns M, Fitzsimmons C, Lindgren G, Sandberg K, Baumung R, Vetterlein M, Strömberg S, Grabherr M, Wade C, Lindblad-Toh K, Pontén F, Heldin CH, Sölkner J, Andersson L. A cis-acting regulatory mutation causes premature hair graying and susceptibility to melanoma in the horse. Nat Genet 2008;40(8):1004–1009.
                        doi: 10.1038/ng.185pubmed: 18641652google scholar: lookup
                      18. Sundstrom E, Komisarczuk AZ, Jiang L, Golovko A, Navratilova P, Rinkwitz S, Becker TS, Andersson L. Identification of a melanocyte-specific, microphthalmia-associated transcription factor-dependent regulatory element in the intronic duplication causing hair greying and melanoma in horses. Pigment Cell Melanoma Res 2012;25(1):28–36.
                        doi: 10.1111/j.1755-148X.2011.00902.xpubmed: 21883983google scholar: lookup
                      19. Sundstrom E, Imsland F, Mikko S, Wade C, Sigurdsson S, Pielberg GR, Golovko A, Curik I, Seltenhammer MH, Sölkner J, Lindblad-Toh K, Andersson L. Copy number expansion of the STX17 duplication in melanoma tissue from Grey horses. BMC Genomics 2012;13:365.
                        doi: 10.1186/1471-2164-13-365pmc: PMC3443021pubmed: 22857264google scholar: lookup
                      20. Eskandarpour M, Kiaii S, Zhu C, Castro J, Sakko AJ, Hansson J. Suppression of oncogenic NRAS by RNA interference induces apoptosis of human melanoma cells. Int J Cancer 2005;115(1):65–73.
                        doi: 10.1002/ijc.20873pubmed: 15688405google scholar: lookup
                      21. Johnson JP, Demmer-Dieckmann M, Meo T, Hadam MR, Riethmuller G. Surface antigens of human melanoma cells defined by monoclonal antibodies. I. Biochemical characterization of two antigens found on cell lines and fresh tumours of diverse tissue origin. Eur J Immunol 1981;11(10):825–831.
                        doi: 10.1002/eji.1830111015pubmed: 6895502google scholar: lookup
                      22. Liao SK, Dent PB, McCulloch PB. Characterization of human maligant melanoma cell lines. I. Morphology and growth characteristics in culture. J Natl Cancer Inst 1975;54(5):1037–1044.
                        pubmed: 1127734
                      23. Campagne C, Jule S, Bernex F, Estrada M, Aubin-Houzelstein G, Panthier JJ, Egidy G. RACK1, a clue to the diagnosis of cutaneous melanomas in horses. BMC Vet Res 2012;8:95.
                        doi: 10.1186/1746-6148-8-95pmc: PMC3543212pubmed: 22747534google scholar: lookup
                      24. Seltenhammer MH, Sundström E, Meisslitzer-Ruppitsch C, Cejka P, Kosiuk J, Neumüller J, Almeder M, Majdic O, Steinberger P, Losert UM, Stöckl J, Andersson L, Sölkner J, Vetterlein M, Golovko A. Establishment and characterization of a primary and a metastatic melanoma cell line from Grey horses. In Vitro Cell Dev Biol Anim 2014;50(1):56–65.
                        doi: 10.1007/s11626-013-9678-1pubmed: 23982913google scholar: lookup
                      25. Haase B, Brooks SA, Schlumbaum A, Azor PJ, Bailey E, Alaeddine F, Mevissen M, Burger D, Poncet PA, Rieder S, Leeb T. Allelic heterogeneity at the equine KIT locus in dominant white (W) horses. PLoS Genet 2007;3(11):e195.
                        doi: 10.1371/journal.pgen.0030195pmc: PMC2065884pubmed: 17997609google scholar: lookup
                      26. Marklund S, Moller M, Sandberg K, Andersson L. Close association between sequence polymorphism in the KIT gene and the roan coat color in horses. Mamm Genome 1999;10(3):283–288.
                        doi: 10.1007/s003359900987pubmed: 10051325google scholar: lookup
                      27. Garnett MJ, Marais R. Guilty as charged: B-RAF is a human oncogene. Cancer Cell 2004;6(4):313–319.
                        doi: 10.1016/j.ccr.2004.09.022pubmed: 15488754google scholar: lookup
                      28. 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(5):584–591.
                        doi: 10.1038/nsmb.2022pubmed: 21478863google scholar: lookup
                      29. Calipel A, Mouriaux F, Glotin AL, Malecaze F, Faussat AM, Mascarelli F. Extracellular signal-regulated kinase-dependent proliferation is mediated through the protein kinase A/B-Raf pathway in human uveal melanoma cells. J Biol Chem 2006;281(14):9238–9250.
                        doi: 10.1074/jbc.M600228200pubmed: 16452469google scholar: lookup
                      30. Wellbrock C, Marais R. Elevated expression of MITF counteracts B-RAF-stimulated melanocyte and melanoma cell proliferation. J Cell Biol 2005;170(5):703–708.
                        doi: 10.1083/jcb.200505059pmc: PMC2171350pubmed: 16129781google scholar: lookup
                      31. Cohen C, Zavala-Pompa A, Sequeira JH, Shoji M, Sexton DG, Cotsonis G, Cerimele F, Govindarajan B, Macaron N, Arbiser JL. Mitogen-actived map kinase activation is an early event in melanoma progression. Clin Cancer Res 2002;8(12):3728–3733.
                        pubmed: 12473582
                      32. Govindarajan B, Bai X, Cohen C, Zhong H, Kilroy S, Louis G, Moses M, Arbiser JL. Malignant transformation of melanocytes to melanoma by constitutive activation of mitogen-activated protein kinase kinase (MAPKK) signaling. J Biol Chem 2003;278(11):9790–9795.
                        doi: 10.1074/jbc.M212929200pubmed: 12514183google scholar: lookup
                      33. The pre-publication history for this paper can be accessed here: http://www.biomedcentral.com/1471-2407/14/857/prepub

                      Citations

                      This article has been cited 16 times.
                      1. Tesena P, Vinijkumthorn R, Kingkaw A, Yanyongsirikarn P, Phasuk K, Ploypetch S, Phaonakrop N, Roytrakul S, Vongsangnak W, Prapaiwan N. Probing Wnt pathway and functional signal in equine melanocytic neoplasms through quantitative proteomics and immunohistochemistry. BMC Vet Res 2025 Aug 7;21(1):509.
                        doi: 10.1186/s12917-025-04956-wpubmed: 40775356google scholar: lookup
                      2. Kurhaluk N, Tkaczenko H. Recent Issues in the Development and Application of Targeted Therapies with Respect to Individual Animal Variability. Animals (Basel) 2025 Feb 6;15(3).
                        doi: 10.3390/ani15030444pubmed: 39943214google scholar: lookup
                      3. Andersson L. White horses - non-coding sequences drive premature hair greying and predisposition to melanoma. Ups J Med Sci 2024;129.
                        doi: 10.48101/ujms.v129.10626pubmed: 38571883google scholar: lookup
                      4. Gao Y, Packeiser EM, Wendt S, Sekora A, Cavalleri JV, Pratscher B, Alammar M, Hühns M, Brenig B, Junghanss C, Nolte I, Murua Escobar H. Cross-Species Comparison of the Pan-RAF Inhibitor LY3009120's Anti-Tumor Effects in Equine, Canine, and Human Malignant Melanoma Cell Lines. Genes (Basel) 2024 Feb 3;15(2).
                        doi: 10.3390/genes15020202pubmed: 38397192google scholar: lookup
                      5. Kuo FC, Tsai HY, Cheng BL, Tsai KJ, Chen PC, Huang YB, Liu CJ, Wu DC, Wu MC, Huang B, Lin MW. Endothelial Mitochondria Transfer to Melanoma Induces M2-Type Macrophage Polarization and Promotes Tumor Growth by the Nrf2/HO-1-Mediated Pathway. Int J Mol Sci 2024 Feb 3;25(3).
                        doi: 10.3390/ijms25031857pubmed: 38339136google scholar: lookup
                      6. Pimenta J, Prada J, Cotovio M. Equine Melanocytic Tumors: A Narrative Review. Animals (Basel) 2023 Jan 10;13(2).
                        doi: 10.3390/ani13020247pubmed: 36670786google scholar: lookup
                      7. Yi Z, Gao Y, Yu F, Zhu Y, Liu H, Li J, Murua Escobar H. Interventions for treatment of cutaneous melanoma in horses: a structured literature review. Vet Res Commun 2023 Jun;47(2):347-360.
                        doi: 10.1007/s11259-022-10023-8pubmed: 36329228google scholar: lookup
                      8. Gregg RK. Model Systems for the Study of Malignant Melanoma. Methods Mol Biol 2021;2265:1-21.
                        doi: 10.1007/978-1-0716-1205-7_1pubmed: 33704702google scholar: lookup
                      9. Ganbaatar O, Konnai S, Okagawa T, Nojima Y, Maekawa N, Minato E, Kobayashi A, Ando R, Sasaki N, Miyakoshi D, Ichii O, Kato Y, Suzuki Y, Murata S, Ohashi K. PD-L1 expression in equine malignant melanoma and functional effects of PD-L1 blockade. PLoS One 2020;15(11):e0234218.
                        doi: 10.1371/journal.pone.0234218pubmed: 33216754google scholar: lookup
                      10. Minnoye L, Taskiran II, Mauduit D, Fazio M, Van Aerschot L, Hulselmans G, Christiaens V, Makhzami S, Seltenhammer M, Karras P, Primot A, Cadieu E, van Rooijen E, Marine JC, Egidy G, Ghanem GE, Zon L, Wouters J, Aerts S. Cross-species analysis of enhancer logic using deep learning. Genome Res 2020 Dec;30(12):1815-1834.
                        doi: 10.1101/gr.260844.120pubmed: 32732264google scholar: lookup
                      11. van der Weyden L, Brenn T, Patton EE, Wood GA, Adams DJ. Spontaneously occurring melanoma in animals and their relevance to human melanoma. J Pathol 2020 Sep;252(1):4-21.
                        doi: 10.1002/path.5505pubmed: 32652526google scholar: lookup
                      12. Liu G, Li C, Zhen H, Zhang Z, Sha Y. Identification of prognostic gene biomarkers for metastatic skin cancer using data mining. Biomed Rep 2020 Jul;13(1):22-30.
                        doi: 10.3892/br.2020.1307pubmed: 32494360google scholar: lookup
                      13. Qi JC, Xue WY, Zhang YP, Qu CB, Lu BS, Yin YW, Liu KL, Wang DB, Li W, Zhao ZM. Cholinergic α5 nicotinic receptor is involved in the proliferation and invasion of human prostate cancer cells. Oncol Rep 2020 Jan;43(1):159-168.
                        doi: 10.3892/or.2019.7411pubmed: 31789411google scholar: lookup
                      14. Grilz-Seger G, Druml T, Neuditschko M, Dobretsberger M, Horna M, Brem G. High-resolution population structure and runs of homozygosity reveal the genetic architecture of complex traits in the Lipizzan horse. BMC Genomics 2019 Mar 5;20(1):174.
                        doi: 10.1186/s12864-019-5564-xpubmed: 30836959google scholar: lookup
                      15. Bourneuf E. The MeLiM Minipig: An Original Spontaneous Model to Explore Cutaneous Melanoma Genetic Basis. Front Genet 2017;8:146.
                        doi: 10.3389/fgene.2017.00146pubmed: 29081790google scholar: lookup
                      16. van der Weyden L, Patton EE, Wood GA, Foote AK, Brenn T, Arends MJ, Adams DJ. Cross-species models of human melanoma. J Pathol 2016 Jan;238(2):152-65.
                        doi: 10.1002/path.4632pubmed: 26354726google scholar: lookup
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