Molecular diagnosis of clear cell sarcoma: Detection of EWS-ATF1 and MITF-M transcripts and histopathological and ultrastructural analysis of 12 cases Journal Article


Authors: Antonescu, C. R.; Tschernyavsky, S. J.; Woodruff, J. M.; Jungbluth, A. A.; Brennan, M. F.; Ladanyi, M.
Article Title: Molecular diagnosis of clear cell sarcoma: Detection of EWS-ATF1 and MITF-M transcripts and histopathological and ultrastructural analysis of 12 cases
Abstract: Clear cell sarcoma (CCS), also known as melanoma of soft parts, is an uncommon deep soft tissue tumor presenting typically in the lower extremities of young adults. Previous cytogenetic studies have established the specificity of the recurrent t(12;22Xq13;q12), resulting in a EWS-ATF1 fusion, for CCS. The prevalence of the EWS-ATF1 fusion in CCS remains unclear, since most genetically confirmed CCS have been reported as isolated cytogenetic or molecular diagnostic case reports. We therefore studied histologically confirmed CCS from 12 patients for the presence of EWS-ATF1 by reverse-transcriptase polymerase chain reaction (RT-PCR), using RNA extracted from either frozen (four cases) or formalin-fixed paraffin-embedded (eight cases) material. All primary tumors were located in the deep soft tissues of the extremities. Histologically, 10 cases had a typical epithelioid nested appearance. Most or all cases showed immunostaining for HMB45 (12 of 12), S-100 protein (10 of 12), and MTTF (12 of 12). Ultrastructural analysis showed melanosomes in six of seven cases. The presence of an EWS-ATF1 fusion transcript was identified by RT-PCR in 11 of 12 cases (91%), all of which showed the same fusion transcript structure, namely the previously described in-frame fusion of EWS exon 8 to ATF1 codon 65. RT-PCR analysis for the melanocyte-specific splice form of the MITF transcript was positive in all cases tested (4 of 4). These data confirm that EWS-ATF1 detection can be used as a highly sensitive diagnostic test for CCS and that CCS expresses the melanocyte-specific form of the MITF transcript, further supporting its genuine melanocytic differentiation. Copyright © American Society for Investigative Pathology and the Association for Molecular Pathology.
Keywords: immunohistochemistry; adult; clinical article; controlled study; human tissue; aged; middle aged; dna binding protein; oncoprotein; gene translocation; human cell; genetics; dna-binding proteins; histopathology; prospective study; prospective studies; genetic analysis; chromosome 12; metabolism; reverse transcription polymerase chain reaction; melanocyte; melanocytes; prevalence; neoplasm proteins; transcription factor; genetic transcription; cell differentiation; pathology; transcription factors; sarcoma; rna; reverse transcriptase polymerase chain reaction; tumor protein; gene fusion; oncogene proteins, fusion; translocation, genetic; protein s 100; dna primers; primer dna; codon; microphthalmia associated transcription factor; rna binding protein ews; epithelioid cell; chromosome analysis; rna, neoplasm; ultrastructure; tumor diagnosis; ews atf1 fusion protein, human; ews-atf1 fusion protein, human; clear cell sarcoma; rna extraction; sarcoma, clear cell; s100 proteins; chromosome 22; chromosomes, human, pair 22; activating transcription factor 1; melanosome; chromosomes, human, pair 12; humans; human; male; female; article; melanoma specific antigens; melanoma-specific antigens; mitf protein, human; s100a1 protein; microphthalmia-associated transcription factor
Journal Title: Journal of Molecular Diagnostics
Volume: 4
Issue: 1
ISSN: 1525-1578
Publisher: Elsevier Science, Inc.  
Date Published: 2002-02-01
Start Page: 44
End Page: 52
Language: English
PUBMED: 11826187
PROVIDER: scopus
PMCID: PMC1906974
DOI: 10.1016/S1525-1578(10)60679-4
DOI/URL:
Notes: Export Date: 14 November 2014 -- Source: Scopus
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MSK Authors
  1. Murray F Brennan
    782 Brennan
  2. Cristina R Antonescu
    612 Antonescu
  3. Marc Ladanyi
    879 Ladanyi
  4. Achim Jungbluth
    341 Jungbluth
  5. James M Woodruff
    138 Woodruff