Molecular detection of EWS-FLI1 chimeric transcripts in Ewing family tumors by nested reverse transcription-polymerase chain reaction: application to archival paraffin-embedded tumor tissues.

Abstract

Chromosomal translocations generating unique chimeric genes are highly characteristic of specific sarcomas, and their use as diagnostic markers has been suggested. From a diagnostic pathologic point of view, detection of such cytogenetic or molecular aberrations applicable to routinely processed archival tissue specimens is considered a powerful tool for tumor diagnosis. To assess the feasibility and reliability of the molecular detection of the transcript originating from the chimeric gene in paraffin-embedded tumor specimens, we performed a nested reverse transcription-polymerase chain reaction (RT-PCR)-based assay to detect the EWS-FLI1 chimeric message in a series of Ewing family tumors. Of 24 paraffin-embedded tumor specimens from 23 cases analyzed, the chimeric message was detectable in 20 (83%) specimens from 20 cases (87%) by this nested RT-PCR assay, whereas none of 7 small round cell tumors not from this family (3 alveolar rhabdomyosarcomas, 2 neuroblastomas, 2 malignant lymphomas) showed detectable chimeric messages. In the sequence analysis of the PCR products, the amplified chimeric messages contained the junctions between exon 7 of the EWS gene and any one of exons 5, 6 and 8 of the FLI1 gene. The detection process was usually completed within 3 days, except for the subseqent sequence analysis. Our results endorse the use of this molecular assay as an ancillary technique in the diagnosis of Ewing family tumors using paraffin-embedded material.

• Authors: M Hisaoka, S Tsuji, Y Morimitsu, H Hashimoto, S Shimajiri, S Komiya, M Ushijima
• Journal: APMIS : acta pathologica, microbiologica, et immunologica Scandinavica (APMIS) Vol. 107 Issue 6 Pg. 577-84 (Jun 1999) ISSN: 0903-4641 [Print] Denmark
• PMID: 10379685 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• EWS-FLI fusion protein
• Oncogene Proteins, Fusion
• Proto-Oncogene Protein c-fli-1
• RNA-Binding Protein EWS
• Transcription Factors
• Paraffin

Topics

• Adolescent
• Adult
• Aged
• Bone Neoplasms (genetics)
• Child
• Child, Preschool
• Chromosomes, Human, Pair 11
• Chromosomes, Human, Pair 22
• Exons
• Female
• Humans
• In Situ Hybridization, Fluorescence
• Infant
• Male
• Middle Aged
• Oncogene Proteins, Fusion (analysis, genetics)
• Paraffin
• Polymerase Chain Reaction
• Proto-Oncogene Protein c-fli-1
• RNA-Binding Protein EWS
• Sarcoma, Ewing (genetics)
• Sequence Analysis, DNA
• Tissue Embedding
• Transcription Factors (analysis, genetics)
• Translocation, Genetic