Reverse transcriptase polymerase chain reaction for detecting Ewing's sarcoma in archival fine needle aspiration biopsies.
OBJECTIVE
Previous studies on pediatric soft tissue sarcomas have demonstrated a chromosomal 11;22 (q24;q12) translocation in Ewing's sarcoma (ES) and peripheral primitive neuroectodermal tumors that appears to be a unique oncogenic marker. To investigate the usefulness of reverse transcriptase polymerase chain reaction (RT-PCR) as an ancillary method for cytodiagnosis, we tested archival aspirates derived from ES patients to establish whether any beta-actin RNA expression or tumor-specific EWS/FLI-1 gene translocation had occurred.


METHODS
Sixteen skeletal specimens aspirated 10-15 years earlier from patients with cytologic and histologic diagnoses of ES were prepared for PCR. The amplification products were sequenced.


RESULTS
Amplifiable RNA was detected in smears from 12 patients by beta-actin RT-PCR. Seven aspirates from beta-actin-positive patients showed ES-specific genomic translocation (58%). Sequence analysis of the resulting PCR fragments revealed EWS/FLI-1 fusion transcriptions of varying length.


CONCLUSIONS
When RNA was retrievable, RT-PCR applied to routinely stained aspirate smears was a highly specific method in differential diagnosis of ES.
Schlott
T
T
Department of Pathology, Georg-August-University, Göttingen, Germany.
Nagel
H
H
Ruschenburg
I
I
Reimer
S
S
Droese
M
M
D001483:Base Sequence; D001707:Biopsy, Needle; D001859:Bone Neoplasms / Q000175:diagnosis* / Q000235:genetics; D002880:Chromosomes, Human, Pair 11; D002892:Chromosomes, Human, Pair 22; D003581:Cytodiagnosis / Q000379:methods*; D004268:DNA-Binding Proteins / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D051789:Proto-Oncogene Protein c-fli-1; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger / Q000032:analysis; D012512:Sarcoma, Ewing / Q000175:diagnosis* / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics*; D014178:Translocation, Genetic
D004268:DNA-Binding Proteins; D051789:Proto-Oncogene Protein c-fli-1; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D015534:Trans-Activators