Abstract |
Most Ewing tumors (ET), including Ewing sarcomas, peripheral primitive neuroectodermal tumors ( PNET), and Askin's tumors, can be defined according to the specific chromosomal translocations t(11;22)(q24;q12) (EWS-FLI-1) or t(21;22)(q21;q12) (EWS-ERG). Detection of the chimeric RNA transcripts by reverse transcriptase-polymerase chain reaction (RT-PCR) has greatly facilitated the diagnosis of ET. Because of variable chromosomal breakpoint locations, however, the EWS gene fusions with FLI-1 and ERG genes are highly heterogenous, resulting in different sizes of the amplification products. To improve the diagnostic usefulness of the RT-PCR assay, we have developed an assay to detect chimeric mRNA transcripts by nested RT-PCR, followed by digestion of the PCR fragments with three different restriction endonucleases. This allows confirmation of the specificity of the PCR product and provides a rapid method to determine the combination of exons present in a transcript. In the 12 Ewing tumors tested, five different exon combinations were detected. In nine repeat biopsies of four patients, the case-specific translocation remained unchanged. One additional central PNET had no ET-specific translocation. In conclusion, the suggested combination of RT-PCR and restriction analysis of the PCR products allows a rapid and specific determination of ET-specific translocations.
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Authors | V S Meier, T Kühne, G Jundt, F Gudat |
Journal | Diagnostic molecular pathology : the American journal of surgical pathology, part B
(Diagn Mol Pathol)
Vol. 7
Issue 1
Pg. 29-35
(Feb 1998)
ISSN: 1052-9551 [Print] United States |
PMID | 9646032
(Publication Type: Journal Article)
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Chemical References |
- DNA-Binding Proteins
- ERG protein, human
- EWS-FLI fusion protein
- Oncogene Proteins
- Oncogene Proteins, Fusion
- Proto-Oncogene Protein c-fli-1
- RNA-Binding Protein EWS
- Trans-Activators
- Transcription Factors
- Transcriptional Regulator ERG
- RNA-Directed DNA Polymerase
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Topics |
- Adolescent
- Adult
- Child
- Chromosomes, Human, Pair 11
- Chromosomes, Human, Pair 21
- Chromosomes, Human, Pair 22
- DNA-Binding Proteins
- Exons
(genetics)
- Female
- Humans
- Male
- Oncogene Proteins
(genetics)
- Oncogene Proteins, Fusion
(genetics)
- Polymerase Chain Reaction
- Proto-Oncogene Protein c-fli-1
- RNA-Binding Protein EWS
- RNA-Directed DNA Polymerase
- Sarcoma, Ewing
(diagnosis, genetics)
- Trans-Activators
- Transcription Factors
(genetics)
- Transcription, Genetic
- Transcriptional Regulator ERG
- Translocation, Genetic
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