Skip to main content

Fluorescence In Situ Hybridization (FISH) Detection of Chromosomal 12p Anomalies in Testicular Germ Cell Tumors

  • Protocol
  • First Online:
Testicular Germ Cell Tumors


Gains of genetic material or internal rearrangements of chromosome 12p, including 12p overrepresentation or isochromosome 12p [i(12p)], are observed in virtually all germ cell tumors (GCT), in all histologic subtypes, and from various body locations. The chromosomal region involved in these alterations contains the growth and survival promoting oncogene KRAS (12p12.1). Gains or rearrangements of 12p characterize GCT from in situ to chemoresistant stages. Fluorescence in situ hybridization (FISH) detection of chromosome 12p anomalies is a sensitive and specific test for the diagnosis of germ cell tumors. Here we provide a detailed protocol for FISH detection of isochromosome 12p and chromosome 12p overrepresentation. The method is helpful for diagnosis of germ cell origin, and for selection of patients who may benefit from cisplatin-based chemotherapy.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others


  1. Ghazarian AA, Kelly SP, Altekruse SF, Rosenberg PS, McGlynn KA (2017) Future of testicular germ cell tumor incidence in the United States: forecast through 2026. Cancer 123:2320–2328

    Article  CAS  Google Scholar 

  2. Fitzmaurice C, Allen C, Barber RM, Barregard L, Bhutta ZA, Brenner H et al (2017) Global, regional, and national cancer incidence, mortality, years of life lost, years lived with disability, and disability-adjusted life-years for 32 cancer groups, 1990 to 2015: a systematic analysis for the global burden of disease study. JAMA Oncol 3:524–548

    Article  Google Scholar 

  3. Chovanec M, Hanna N, Cary KC, Einhorn L, Albany C (2016) Management of stage I testicular germ cell tumours. Nat Rev Urol 13:663–673

    Article  CAS  Google Scholar 

  4. Ruf CG, Isbarn H, Wagner W, Fisch M, Matthies C, Dieckmann KP (2014) Changes in epidemiologic features of testicular germ cell cancer: age at diagnosis and relative frequency of seminoma are constantly and significantly increasing. Urol Oncol 32:33.e1–36.e1

    Article  Google Scholar 

  5. Cheng L, Lyu B, Roth LM (2017) Perspectives on testicular germ cell neoplasms. Hum Pathol 59:10–25

    Article  CAS  Google Scholar 

  6. Roth LM, Cheng L (2016) Mixed germ cell-sex cord stromal tumor of the testis with an intratubular component: a problem in differential diagnosis. Hum Pathol 51:51–56

    Article  Google Scholar 

  7. Taylor-Weiner A, Zack T, O’Donnell E, Guerriero JL, Bernard B, Reddy A et al (2016) Genomic evolution and chemoresistance in germ-cell tumours. Nature 540:114–118

    Article  CAS  Google Scholar 

  8. Litchfield K, Summersgill B, Yost S, Sultana R, Labreche K, Dudakia D et al (2015) Whole-exome sequencing reveals the mutational spectrum of testicular germ cell tumours. Nat Commun 6:5973

    Article  CAS  Google Scholar 

  9. Litchfield K, Levy M, Huddart RA, Shipley J, Turnbull C (2016) The genomic landscape of testicular germ cell tumours: from susceptibility to treatment. Nat Rev Urol 13:409–419

    Article  CAS  Google Scholar 

  10. Dmitrovsky E, Murty VV, Moy D, Miller WH Jr, Nanus D, Albino AP et al (1990) Isochromosome 12p in non-seminoma cell lines: karyologic amplification of c-ki-ras2 without point-mutational activation. Oncogene 5:543–548

    CAS  PubMed  Google Scholar 

  11. Suijkerbuijk RF, Looijenga L, de Jong B, Oosterhuis JW, Cassiman JJ, Geurts van Kessel A (1992) Verification of isochromosome 12p and identification of other chromosome 12 aberrations in gonadal and extragonadal human germ cell tumors by bicolor double fluorescence in situ hybridization. Cancer Genet Cytogenet 63:8–16

    Article  CAS  Google Scholar 

  12. Mostert MM, van de Pol M, van Echten J, Olde Weghuis D, Geurts van Kessel A, Oosterhuis JW et al (1996) Fluorescence in situ hybridization-based approaches for detection of 12p overrepresentation, in particular i(12p), in cell lines of human testicular germ cell tumors of adults. Cancer Genet Cytogenet 87:95–102

    Article  CAS  Google Scholar 

  13. Cheng L, Zhang S, Eble JN, Beck SD, Foster RS, Wang M et al (2012) Molecular genetic evidence supporting the neoplastic nature of fibrous stroma in testicular teratoma. Mod Pathol 25:1432–1438

    Article  CAS  Google Scholar 

  14. Emerson RE, Kao CS, Eble JN, Grignon DJ, Wang M, Zhang S et al (2017) Evidence of a dual histogenetic pathway of sacrococcygeal teratomas. Histopathology 70:290–300

    Article  Google Scholar 

  15. Cheng L, Zhang D (2008) Molecular genetic pathology. Springer, New York

    Book  Google Scholar 

  16. Cheng L, Zhang S, Wang M, Davidson DD, Morton MJ, Huang J et al (2007) Molecular genetic evidence supporting the neoplastic nature of stromal cells in “fibrosis” after chemotherapy for testicular germ cell tumors. J Pathol 213:65–71

    Article  CAS  Google Scholar 

  17. Poulos C, Cheng L, Zhang S, Gersell DJ, Ulbright TM (2006) Analysis of ovarian teratomas for isochromosome 12p: evidence supporting a dual histogenetic pathway for teratomatous elements. Mod Pathol 19:766–771

    Article  CAS  Google Scholar 

  18. Cheng L, Zhang S, MacLennan GT, Poulos CK, Sung MT, Beck SD et al (2006) Interphase fluorescence in situ hybridization analysis of chromosome 12p abnormalities is useful for distinguishing epidermoid cysts of the testis from pure mature teratoma. Clin Cancer Res 12:5668–5672

    Article  CAS  Google Scholar 

  19. Cossu-Rocca P, Zhang S, Roth LM, Eble JN, Zheng W, Karim FW et al (2006) Chromosome 12p abnormalities in dysgerminoma of the ovary: a FISH analysis. Mod Pathol 19:611–615

    Article  CAS  Google Scholar 

  20. Mostert MC, Verkerk AJ, van de Pol M, Heighway J, Marynen P, Rosenberg C et al (1998) Identification of the critical region of 12p over-representation in testicular germ cell tumors of adolescents and adults. Oncogene 16:2617–2627

    Article  CAS  Google Scholar 

  21. Reuter VE (2005) Origins and molecular biology of testicular germ cell tumors. Mod Pathol 18 Suppl 2:S51–S60

    Google Scholar 

  22. Cheng L, Zhang S, Wang L, MacLennan GT, Davidson DD (2017) Fluorescence in situ hybridization in surgical pathology: principles and applications. J Pathol Clin Res 3:73–99

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations


Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Cheng, L. et al. (2021). Fluorescence In Situ Hybridization (FISH) Detection of Chromosomal 12p Anomalies in Testicular Germ Cell Tumors. In: Bagrodia, A., Amatruda, J.F. (eds) Testicular Germ Cell Tumors. Methods in Molecular Biology, vol 2195. Humana, New York, NY.

Download citation

  • DOI:

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0859-3

  • Online ISBN: 978-1-0716-0860-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics