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Advantage of FISH analysis using FKHR probes for an adjunct to diagnosis of rhabdomyosarcomas

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Abstract

Translocations can be detected using fluorescence in situ hybridization (FISH) in formalin-fixed paraffin-embedded tissues. Recently, a commercially available FKHR (13q14) dual-color, break-apart rearrangement probe has been developed. However, the advantages of using this probe have not been reported. This study demonstrated the usefulness of this probe for the clinical diagnosis of rhabdomyosarcomas (RMS). We studied 33 RMS (19 embryonal rhabdomyosarcomas [ERMS], including three sclerosing-type RMS, and 14 alveloar rhabdomyosarcomas [ARMS]). Fluorescence signals were detected for 18 of the 19 (94.7%) ERMS and 13 of the 14 (92.8%) ARMS. A split-signal pattern was detected in 12 of 13 (92.3%) ARMS but was not detected in any of the ERMS, including the three sclerosing-type RMS. Amplification and polyploidy were present in both the ERMS and the ARMS. Our FISH study highlighted the excellent performance of the presently reported commercial break-apart probe for the detection of FKHR gene rearrangements in RMS. Because amplification and polyploidy were detected in both the ERMS and the ARMS, sufficient care should be taken when counting the nuclear signals. No rearrangements of the FKHR gene were found in any of the three sclerosing-type RMS when examined using a FISH assay, supporting the hypothesis that sclerosing RMS can be included as an ERMS.

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Abbreviations

RMS:

rhabdomyosarcomas

ERMS:

embryonal rhabdomyosarcomas

ARMS:

alveolar rhabdomyosarcomas

PRMS:

pleomorphic rhabdomyosarcomas

RT-PCR:

reverse transcriptase–polymerase chain reaction

FFPE:

formalin-fixed paraffin-embedded

FISH:

fluorescence in situ hybridization

H&E:

hematoxylin and eosin,

DAPI II:

4’,6-diamidino-2-phenylindole

CCD:

charge-coupled-device

References

  1. Barr FG (1997) Molecular genetics and pathogenesis of rhabdomyosarcoma. J Pediatr Hematol Oncol 19:483–491

    Article  PubMed  CAS  Google Scholar 

  2. Barr FG, Nauta LE, Davis RJ, Schäfer BW, Nycum LM, Biegel JA (1996) In vivo amplification of the PAX3–FKHR and PAX7–FKHR fusion genes in alveolar rhabdomyosarcoma. Hum Mol Genet 5:15–21

    Article  PubMed  CAS  Google Scholar 

  3. Barr FG, Qualman SJ, Macris MH, Melnyk N, Lawlor ER, Strzelecki DM, Triche TJ, Bridge JA, Sorensen PH (2002) Genetic heterogeneity in the alveolar rhabdomyosarcoma subset without typical gene fusions. Cancer Res 62:4704–4710

    PubMed  CAS  Google Scholar 

  4. Bridge JA, Liu J, Qualman SJ, Suijkerbuijk R, Wenger G, Zhang J, Wan X, Baker KS, Sorensen P, Barr FG (2002) Genomic gains and losses are similar in genetic and histologic subsets of rhabdomyosarcoma, whereas amplification predominates in embryonal with anaplasia and alveolar subtypes. Genes Chromosomes Cancer 33:310–321

    Article  PubMed  CAS  Google Scholar 

  5. Bridge JA, Liu J, Weibolt V, Baker KS, Perry D, Kruger R, Qualman S, Barr F, Sorensen P, Triche T, Suijkerbuijk R (2000) Novel genomic imbalances in embryonal rhabdomyosarcoma revealed by comparative genomic hybridization and fluorescence in situ hybridization: an intergroup rhabdomyosarcoma study. Genes Chromosomes Cancer 27:337–344

    Article  PubMed  CAS  Google Scholar 

  6. Bridge RS, Rajaram V, Dehner LP, Pfeifer JD, Perry A (2006) Molecular diagnosis of Ewing sarcoma/primitive neuroectodermal tumor in routinely processed tissue: a comparison of two FISH strategies and RT-PCR in malignant round cell tumors. Mod Pathol 19:1–8

    Article  PubMed  CAS  Google Scholar 

  7. Cavazzana AO, Schmidt D, Ninfo V, Harms D, Tollot M, Carli M, Treuner J, Betto R, Salviati G (1992) Spindle cell rhabdomyosarcoma. A prognostically favorable variant of rhabdomyosarcoma. Am J Surg Pathol 16:229–235

    PubMed  CAS  Google Scholar 

  8. Croes R, Debiec-Rychter M, Cokelaere K, De Vos R, Hagemeijer A, Sciot R (2005) Adult sclerosing rhabdomyosarcoma: cytogenetic link with embryonal rhabdomyosarcoma. Virchows Arch 446:64–67

    Article  PubMed  Google Scholar 

  9. Davis RJ, D’Cruz CM, Lovell MA, Biegel JA, Barr FG (1994) Fusion of PAX7 to FKHR by the variant t(1;13)(p36;q14) translocation in alveolar rhabdomyosarcoma. Cancer Res 54:2869–2872

    PubMed  CAS  Google Scholar 

  10. Folpe AL, McKenney JK, Bridge JA, Weiss SW (2002) Sclerosing rhabdomyosarcoma in adults: report of four cases of a hyalinizing, matrix-rich variant of rhabdomyosarcoma that may be confused with osteosarcoma, chondrosarcoma, or angiosarcoma. Am J Surg Pathol 26:1175–1183

    Article  PubMed  Google Scholar 

  11. Forus A, Florenes VA, Maelandsmo GM, Meltzer PS, Fodstad O, Myklebost O (1993) Mapping of amplification units in the q13-14 region of chromosome 12 in human sarcomas: some amplica do not include MDM2. Cell Growth Differ 4:1065–1070

    PubMed  CAS  Google Scholar 

  12. Galili N, Davis RJ, Fredericks WJ, Mukhopadhyay S, Rauscher FJ 3rd, Emanuel BS, Rovera G, Barr FG (1993) Fusion of a fork head domain gene to PAX3 in the solid tumour alveolar rhabdomyosarcoma. Nat Genet 5:230–235

    Article  PubMed  CAS  Google Scholar 

  13. Gordon AT, Brinkschmidt C, Anderson J, Coleman N, Dockhorn-Dworniczak B, Pritchard-Jones K, Shipley J (2000) A novel and consistent amplicon at 13q31 associated with alveolar rhabdomyosarcoma. Genes Chromosomes Cancer 28:220–226

    Article  PubMed  CAS  Google Scholar 

  14. Khatib ZA, Matsushime H, Valentine M, Shapiro DN, Sherr CJ, Look AT (1993) Coamplification of the CDK4 gene with MDM2 and GLI in human sarcomas. Cancer Res 53:5535–5541

    PubMed  CAS  Google Scholar 

  15. Kodet R, Newton WA Jr, Hamoudi AB, Asmar L, Jacobs DL, Maurer HM (1993) Childhood rhabdomyosarcoma with anaplastic (pleomorphic) features. A report of the Intergroup Rhabdomyosarcoma Study. Am J Surg Pathol 17:443–453

    PubMed  CAS  Google Scholar 

  16. Koscielniak E, Harms D, Henze G, Jurgens H, Gadner H, Herbst M, Klingebiel T, Schmidt BF, Morgan M, Knietig R, Treuner J (1999) Results of treatment for soft tissue sarcoma in childhood and adolescence: a final report of the German Cooperative Soft Tissue Sarcoma Study CWS-86. J Clin Oncol 17:3706–3719

    PubMed  CAS  Google Scholar 

  17. Leuschner I, Newton WA Jr, Schmidt D, Sachs N, Asmar L, Hamoudi A, Harms D, Maurer HM (1993) Spindle cell variants of embryonal rhabdomyosarcoma in the paratesticular region. A report of the Intergroup Rhabdomyosarcoma Study. Am J Surg Pathol 17:221–230

    Article  PubMed  CAS  Google Scholar 

  18. Mentzel T, Katenkamp D (2000) Sclerosing, pseudovascular rhabdomyosarcoma in adults. Clinicopathological and immunohistochemical analysis of three cases. Virchows Arch 436:305–311

    Article  PubMed  CAS  Google Scholar 

  19. Mhawech-Fauceglia P, Herrmann F, Penetrante R, Beck A, Sait S, Block AM, Odunsi K, Fisher J, Balos L, Cheney RT (2006) Diagnostic utility of FLI-1 monoclonal antibody and dual-colour, break-apart probe fluorescence in situ (FISH) analysis in Ewing’s sarcoma/primitive neuroectodermal tumour (EWS/PNET). A comparative study with CD99 and FLI-1 polyclonal antibodies. Histopathology 49:569–575

    Article  PubMed  CAS  Google Scholar 

  20. Nishio J, Althof PA, Bailey JM, Zhou M, Neff JR, Barr FG, Parham DM, Teot L, Qualman SJ, Bridge JA (2006) Use of a novel FISH assay on paraffin-embedded tissues as an adjunct to diagnosis of alveolar rhabdomyosarcoma. Lab Invest 86:547–556

    PubMed  CAS  Google Scholar 

  21. Pandita A, Zielenska M, Thorner P, Bayani J, Godbout R, Greenberg M, Squire JA (1999) Application of comparative genomic hybridization, spectral karyotyping, and microarray analysis in the identification of subtype-specific patterns of genomic changes in rhabdomyosarcoma. Neoplasia 1:262–275

    Article  PubMed  CAS  Google Scholar 

  22. Parham DM, Barr FG (2002) Embryonal rhabdomyosarcoma. In: Fletcher CDM, Unni KK, Mertens F (eds) WHO classification of tumours, pathology and genetics of tumours of soft tissue and bone. IARC, Lyon, France, pp 146–154

    Google Scholar 

  23. Parham DM, Shapiro DN, Downing JR, Webber BL, Douglass EC (1994) Solid alveolar rhabdomyosarcomas with the t(2;13). Report of two cases with diagnostic implications. Am J Surg Pathol 18:474–478

    Article  PubMed  CAS  Google Scholar 

  24. Patel RM, Downs-Kelly E, Weiss SW, Folpe AL, Tubbs RR, Tuthill RJ, Goldblum JR, Skacel M (2005) Dual-color, break-apart fluorescence in situ hybridization for EWS gene rearrangement distinguishes clear cell sarcoma of soft tissue from malignant melanoma. Mod Pathol 18:1585–1590

    Article  PubMed  CAS  Google Scholar 

  25. Qualman SJ, Coffin CM, Newton WA, Hojo H, Triche TJ, Parham DM, Crist WM (1998) Intergroup Rhabdomyosarcoma Study: update for pathologists. Pediatr Dev Pathol 1:550–561

    Article  PubMed  CAS  Google Scholar 

  26. Roberts WM, Douglass EC, Peiper SC, Houghton PJ, Look AT (1989) Amplification of the gli gene in childhood sarcomas. Cancer Res 49:5407–5413

    PubMed  CAS  Google Scholar 

  27. Sartelet H, Lantuejoul S, Armari-Alla C, Pin I, Delattre O, Brambilla E (1998) Solid alveolar rhabdomyosarcoma of the thorax in a child. Histopathology 32:165–171

    Article  PubMed  CAS  Google Scholar 

  28. Shapiro DN, Jones BG, Shapiro LH, Dias P, Houghton PJ (1994) Antisense-mediated reduction in insulin-like growth factor-I receptor expression suppresses the malignant phenotype of a human alveolar rhabdomyosarcoma. J Clin Invest 94:1235–1242

    Article  PubMed  CAS  Google Scholar 

  29. Sorensen PH, Lynch JC, Qualman SJ, Tirabosco R, Lim JF, Maurer HM, Bridge JA, Crist WM, Triche TJ, Barr FG (2002) PAX3-FKHR and PAX7-FKHR gene fusions are prognostic indicators in alveolar rhabdomyosarcoma: a report from the children’s oncology group. J Clin Oncol 20:2672–2679

    Article  PubMed  CAS  Google Scholar 

  30. Surace C, Panagopoulos I, Palsson E, Rocchi M, Mandahl N, Mertens F (2004) A novel FISH assay for SS18-SSX fusion type in synovial sarcoma. Lab Invest 84:1185–1192

    Article  PubMed  CAS  Google Scholar 

  31. Tsokos M, Webber BL, Parham DM, Wesley RA, Miser A, Miser JS, Etcubanas E, Kinsella T, Grayson J, Glatstein E (1992) Rhabdomyosarcoma. A new classification scheme related to prognosis. Arch Pathol Lab Med 116:847–855

    PubMed  CAS  Google Scholar 

  32. Ventura RA, Martin-Subero JI, Jones M, McParland J, Gesk S, Mason DY, Siebert R (2006) FISH analysis for the detection of lymphoma-associated chromosomal abnormalities in routine paraffin-embedded tissue. J Mol Diagn 8:141–151

    Article  PubMed  CAS  Google Scholar 

  33. Wachtel M, Dettling M, Koscielniak E, Stegmaier S, Treuner J, Simon-Klingenstein K, Buhlmann P, Niggli FK, Schafer BW (2004) Gene expression signatures identify rhabdomyosarcoma subtypes and detect a novel t(2;2)(q35;p23) translocation fusing PAX3 to NCOA1. Cancer Res 64:5539–5545

    Article  PubMed  CAS  Google Scholar 

  34. Weber-Hall S, Anderson J, McManus A, Abe S, Nojima T, Pinkerton R, Pritchard-Jones K, Shipley J (1996) Gains, losses, and amplification of genomic material in rhabdomyosarcoma analyzed by comparative genomic hybridization. Cancer Res 56:3220–3224

    PubMed  CAS  Google Scholar 

  35. Weiss SW, Goldblum JR (2001) Enzinger and Weiss’s soft tissue tumors, 4th edn. Mosby, St. Louis

    Google Scholar 

  36. Yamaguchi U, Hasegawa T, Morimoto Y, Tateishi U, Endo M, Nakatani F, Kawai A, Chuman H, Beppu Y, Endo M, Kurotaki H, Furuta K (2005) A practical approach to the clinical diagnosis of Ewing’s sarcoma/primitive neuroectodermal tumour and other small round cell tumours sharing EWS rearrangement using new fluorescence in situ hybridisation probes for EWSR1 on formalin fixed, paraffin wax embedded tissue. J Clin Pathol 58:1051–1056

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Tadashi Hasegawa.

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Matsumura, T., Yamaguchi, T., Seki, K. et al. Advantage of FISH analysis using FKHR probes for an adjunct to diagnosis of rhabdomyosarcomas. Virchows Arch 452, 251–258 (2008). https://doi.org/10.1007/s00428-007-0554-9

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  • DOI: https://doi.org/10.1007/s00428-007-0554-9

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