Tumor Biology

, Volume 37, Issue 4, pp 4901–4907 | Cite as

Hotspot TERT promoter mutations are rare events in testicular germ cell tumors

  • Flavio Mavignier Cárcano
  • Daniel Onofre Vidal
  • André van Helvoort Lengert
  • Cristovam Scapulatempo Neto
  • Luisa Queiroz
  • Herlander Marques
  • Fátima Baltazar
  • Camila Maria da Silva Martinelli
  • Paula Soares
  • Eduardo Caetano Albino da Silva
  • Luiz Fernando Lopes
  • Rui Manuel Reis
Original Article


The abnormal activation of telomerase, codified by the telomerase reverse transcriptase (TERT) gene, is related to one of cancer hallmarks. Hotspot somatic mutations in the promoter region of TERT, specifically the c.-124:C>T and c.-146:C>T, were recently identified in a range of human cancers and have been associated with a more aggressive behavior. Testicular germ cell tumors frequently exhibit a good prognosis; however, the development of refractory disease is still a clinical challenge. In this study, we aim to evaluate for the first time the presence of the hotspot telomerase reverse transcriptase gene promoter mutations in testicular germ cell tumors. A series of 150 testicular germ cell tumor cases and four germ cell tumor cell lines were evaluated by PCR followed by direct Sanger sequencing and correlated with patient’s clinical pathological features. Additionally, we genotyped the telomerase reverse transcriptase gene promoter single nucleotide polymorphism rs2853669 (T>C) located at −245 position. We observed the presence of the TERT promoter mutation in four patients, one exhibited the c.-124:C>T and three the c.-146:C>T. No association between TERT mutation status and clinicopathological features could be identified. The analysis of the rs2853669 showed that variant C was present in 22.8 % of the cases. In conclusion, we showed for the first time that TERT promoter mutations occur in a small subset (~3 %) of testicular germ cell tumors.


Testicular neoplasms Neoplasms, germ cell, and embryonal TERT protein Mutation Polymorphism, single nucleotide 



The authors acknowledge the support of the Center for Research Support (NAP), Barretos Cancer Hospital, Pio XII Foundation.

Compliance with ethical standards


This work was supported by the Barretos Cancer Hospital internal research funds (PAIP): Project “Microenvironment, metabolism and cancer” that was partially supported by Programa Operacional Regional do Norte (ON.2 – O Novo Norte), under the Quadro de Referência Estratégico Nacional (QREN), and through the Fundo Europeu de Desenvolvimento Regional (FEDER). R.M.R. has a National Counsel of Technological and Scientific Development (CNPq) scholarship.

Conflicts of interest


Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee (register number CAAE 12297713.0.0000.5437) and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. For this type of study, formal consent is not required.


  1. 1.
    Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646–74. doi: 10.1016/j.cell.2011.02.013.CrossRefPubMedGoogle Scholar
  2. 2.
    Huang FW, Hodis E, Xu MJ, Kryukov GV, Chin L, Garraway LA. Highly recurrent TERT promoter mutations in human melanoma. Science. 2013;339(6122):957–9. doi: 10.1126/science.1229259.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Vinagre J, Almeida A, Populo H, Batista R, Lyra J, Pinto V, et al. Frequency of TERT promoter mutations in human cancers. Nat Commun. 2013;4:2185. doi: 10.1038/ncomms3185.CrossRefPubMedGoogle Scholar
  4. 4.
    Killela PJ, Reitman ZJ, Jiao Y, Bettegowda C, Agrawal N, Diaz Jr LA, et al. TERT promoter mutations occur frequently in gliomas and a subset of tumors derived from cells with low rates of self-renewal. Proc Natl Acad Sci U S A. 2013;110(15):6021–6. doi: 10.1073/pnas.1303607110.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Horn S, Figl A, Rachakonda PS, Fischer C, Sucker A, Gast A, et al. TERT promoter mutations in familial and sporadic melanoma. Science. 2013;339(6122):959–61. doi: 10.1126/science.1230062.CrossRefPubMedGoogle Scholar
  6. 6.
    Bell RJ, Rube HT, Kreig A, Mancini A, Fouse SD, Nagarajan RP, et al. Cancer. The transcription factor GABP selectively binds and activates the mutant TERT promoter in cancer. Science. 2015;348(6238):1036–9. doi: 10.1126/science.aab0015.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Park CK, Lee SH, Kim JY, Kim JE, Kim TM, Lee ST, et al. Expression level of hTERT is regulated by somatic mutation and common single nucleotide polymorphism at promoter region in glioblastoma. Oncotarget. 2014;5(10):3399–407.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Rachakonda PS, Hosen I, de Verdier PJ, Fallah M, Heidenreich B, Ryk C, et al. TERT promoter mutations in bladder cancer affect patient survival and disease recurrence through modification by a common polymorphism. Proc Natl Acad Sci U S A. 2013;110(43):17426–31. doi: 10.1073/pnas.1310522110.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Hosen I, Rachakonda PS, Heidenreich B, Sitaram RT, Ljungberg B, Roos G, et al. TERT promoter mutations in clear cell renal cell carcinoma. Int J Cancer. 2014. doi: 10.1002/ijc.29279.PubMedGoogle Scholar
  10. 10.
    Bray F, Ferlay J, Devesa SS, McGlynn KA, Moller H. Interpreting the international trends in testicular seminoma and nonseminoma incidence. Nat Clin Pract Urol. 2006;3(10):532–43. doi: 10.1038/ncpuro0606.CrossRefPubMedGoogle Scholar
  11. 11.
    Einhorn LH. Treatment of testicular cancer: a new and improved model. J Clin Oncol: Off J Am Soc Clin Oncol. 1990;8(11):1777–81.CrossRefGoogle Scholar
  12. 12.
    Lorch A, Beyer J, Bascoul-Mollevi C, Kramar A, Einhorn LH, Necchi A, et al. Prognostic factors in patients with metastatic germ cell tumors who experienced treatment failure with cisplatin-based first-line chemotherapy. J Clin Oncol: Off J Am Soc Clin Oncol. 2010;28(33):4906–11. doi: 10.1200/JCO.2009.26.8128.CrossRefGoogle Scholar
  13. 13.
    Mead GM, Cullen MH, Huddart R, Harper P, Rustin GJ, Cook PA, et al. A phase II trial of TIP (paclitaxel, ifosfamide and cisplatin) given as second-line (post-BEP) salvage chemotherapy for patients with metastatic germ cell cancer: a medical research council trial. Br J Cancer. 2005;93(2):178–84. doi: 10.1038/sj.bjc.6602682.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Albanell J, Bosl GJ, Reuter VE, Engelhardt M, Franco S, Moore MA, et al. Telomerase activity in germ cell cancers and mature teratomas. J Natl Cancer Inst. 1999;91(15):1321–6.CrossRefPubMedGoogle Scholar
  15. 15.
    Turnbull C, Rapley EA, Seal S, Pernet D, Renwick A, Hughes D, et al. Variants near DMRT1, TERT and ATF7IP are associated with testicular germ cell cancer. Nat Genet. 2010;42(7):604–7. doi: 10.1038/ng.607.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Dirks WG, Faehnrich S, Estella IA, Drexler HG. Short tandem repeat DNA typing provides an international reference standard for authentication of human cell lines. ALTEX. 2005;22(2):103–9.PubMedGoogle Scholar
  17. 17.
    Martinho O, Longatto-Filho A, Lambros MB, Martins A, Pinheiro C, Silva A, et al. Expression, mutation and copy number analysis of platelet-derived growth factor receptor A (PDGFRA) and its ligand PDGFA in gliomas. Br J Cancer. 2009;101(6):973–82. doi: 10.1038/sj.bjc.6605225.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Campanella NC, Celestino R, Pestana A, Scapulatempo-Neto C, de Oliveira AT, Brito MJ, et al. Low frequency of TERT promoter mutations in gastrointestinal stromal tumors (GISTs). Eur J Hum Genet: EJHG. 2014. doi: 10.1038/ejhg.2014.195.PubMedCentralGoogle Scholar
  19. 19.
    Campanella NC, Penna V, Abrahao-Machado LF, Cruvinel-Carloni A, Ribeiro G, Soares P, et al. TERT promoter mutations in soft tissue sarcomas. Int J Biol Markers. 2015. doi: 10.5301/jbm.5000168.Google Scholar
  20. 20.
    IGCCCG. International Germ Cell Consensus Classification: a prognostic factor-based staging system for metastatic germ cell cancers. Int Germ Cell Cancer Collab Group J Clin Oncol: Off J Am Soc Clin Oncol. 1997;15(2):594–603.Google Scholar
  21. 21.
    Nault JC, Mallet M, Pilati C, Calderaro J, Bioulac-Sage P, Laurent C, et al. High frequency of telomerase reverse-transcriptase promoter somatic mutations in hepatocellular carcinoma and preneoplastic lesions. Nat Commun. 2013;4:2218. doi: 10.1038/ncomms3218.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Heidenreich B, Rachakonda PS, Hemminki K, Kumar R. TERT promoter mutations in cancer development. Curr Opin Genet Dev. 2014;24c:30–7. doi: 10.1016/j.gde.2013.11.005.CrossRefGoogle Scholar
  23. 23.
    Melo M, da Rocha AG, Vinagre J, Batista R, Peixoto J, Tavares C, et al. TERT promoter mutations are a major indicator of poor outcome in differentiated thyroid carcinomas. J Clin Endocrinol Metab. 2014;99(5):E754–65. doi: 10.1210/jc.2013-3734.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Populo H, Boaventura P, Vinagre J, Batista R, Mendes A, Caldas R, et al. TERT promoter mutations in skin cancer: the effects of sun exposure and x-irradiation. J Invest Dermatol. 2014. doi: 10.1038/jid.2014.163.PubMedGoogle Scholar
  25. 25.
    Honecker F, Wermann H, Mayer F, Gillis AJ, Stoop H, van Gurp RJ, et al. Microsatellite instability, mismatch repair deficiency, and BRAF mutation in treatment-resistant germ cell tumors. J Clin Oncol: Off J Am Soc Clin Oncol. 2009;27(13):2129–36. doi: 10.1200/JCO.2008.18.8623.CrossRefGoogle Scholar
  26. 26.
    Sommerer F, Hengge UR, Markwarth A, Vomschloss S, Stolzenburg JU, Wittekind C, et al. Mutations of BRAF and RAS are rare events in germ cell tumours. Int J Cancer. 2005;113(2):329–35. doi: 10.1002/ijc.20567.CrossRefPubMedGoogle Scholar
  27. 27.
    Vinagre J, Pinto V, Celestino R, Reis M, Populo H, Boaventura P, et al. Telomerase promoter mutations in cancer: an emerging molecular biomarker? Virchows Arch: Int J Pathol. 2014;465(2):119–33. doi: 10.1007/s00428-014-1608-4.CrossRefGoogle Scholar
  28. 28.
    Abecasis GR, Auton A, Brooks LD, DePristo MA, Durbin RM, Handsaker RE, et al. An integrated map of genetic variation from 1,092 human genomes. Nature. 2012;491(7422):56–65. doi: 10.1038/nature11632.CrossRefPubMedGoogle Scholar
  29. 29.
    Dolci S, Levati L, Pellegrini M, Faraoni I, Graziani G, Di Carlo A, et al. Stem cell factor activates telomerase in mouse mitotic spermatogonia and in primordial germ cells. J Cell Sci. 2002;115(Pt 8):1643–9.PubMedGoogle Scholar
  30. 30.
    Schrader M, Burger AM, Muller M, Krause H, Straub B, Smith GL, et al. Quantification of human telomerase reverse transcriptase mRNA in testicular germ cell tumors by quantitative fluorescence real-time RT-PCR. Oncol Rep. 2002;9(5):1097–105.PubMedGoogle Scholar
  31. 31.
    Nowak R, Sikora K, Pietas A, Skoneczna I, Chrapusta SJ. Germ cell-like telomeric length homeostasis in nonseminomatous testicular germ cell tumors. Oncogene. 2000;19(35):4075–8. doi: 10.1038/sj.onc.1203746.CrossRefPubMedGoogle Scholar
  32. 32.
    Cesare AJ, Reddel RR. Alternative lengthening of telomeres: models, mechanisms and implications. Nat Rev Genet. 2010;11(5):319–30. doi: 10.1038/nrg2763.CrossRefPubMedGoogle Scholar
  33. 33.
    Ries LAG YJ, Keel GE, Eisner MP, Lin YD, Horner M-J (editors). SEER survival monograph: cancer survival among adults: U.S. SEER Program, 1988–2001, Patient and Tumor Characteristics. National Cancer Institute, SEER Program, NIH Pub. No. 07–6215, Bethesda, MD, 2007; p165-70. 2007.Google Scholar
  34. 34.
    Lopes LF, Macedo CR, Pontes EM, Dos Santos AS, Mastellaro MJ, Melaragno R, et al. Cisplatin and etoposide in childhood germ cell tumor: Brazilian pediatric oncology society protocol GCT-91. J Clin Oncol: Off J Am Soc Clin Oncol. 2009;27(8):1297–303. doi: 10.1200/JCO.2008.16.4202.CrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Flavio Mavignier Cárcano
    • 1
    • 2
  • Daniel Onofre Vidal
    • 3
    • 4
  • André van Helvoort Lengert
    • 3
    • 4
  • Cristovam Scapulatempo Neto
    • 3
    • 5
  • Luisa Queiroz
    • 6
  • Herlander Marques
    • 6
  • Fátima Baltazar
    • 7
    • 8
  • Camila Maria da Silva Martinelli
    • 3
  • Paula Soares
    • 9
    • 10
    • 11
  • Eduardo Caetano Albino da Silva
    • 5
  • Luiz Fernando Lopes
    • 3
    • 4
  • Rui Manuel Reis
    • 3
    • 7
    • 8
  1. 1.Department of Clinical OncologyBarretos Cancer HospitalBarretosBrazil
  2. 2.Barretos School of Health Sciences, Dr. Paulo Prata–FACISBBarretosBrazil
  3. 3.Molecular Oncology Research CenterBarretos Cancer HospitalBarretosBrazil
  4. 4.Barretos Children’s Cancer HospitalBarretosBrazil
  5. 5.Department of PathologyBarretos Cancer HospitalBarretosBrazil
  6. 6.Department of Medical OncologyHospital de BragaBragaPortugal
  7. 7.Life and Health Sciences Research Institute (ICVS), Health Sciences SchoolUniversity of MinhoBragaPortugal
  8. 8.ICVS/3B’s-PT Government Associate LaboratoryBraga/GuimarãesPortugal
  9. 9.Institute of Molecular Pathology and Immunology of the University of Porto–IPATIMUPPortoPortugal
  10. 10.Instituto de Investigação e Inovação em SaúdeUniversidade do PortoPortoPortugal
  11. 11.Medical FacultyUniversity of PortoPortoPortugal

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