[Application of fluorescence in-situ hybridization and reverse transcription-polymerase chain reaction in molecular diagnosis of Ewing's sarcoma and primitive neuroectodermal tumor].
OBJECTIVE
To detect tumor specific chromosome translocations and associated fusion transcripts in paraffin-embedded tissue by interphase fluorescence in-situ hybridization (FISH) and reverse transcription polymerase chain reaction (RT-PCR), respectively, and to evaluate their diagnostic values for Ewing's sarcoma/primitive neuroectodermal tumor (ES/PNET).


METHODS
Nuclei of the tumor cells and total RNA were extracted from 10 cases of ES/PNET. Interphase FISH was utilized to analyze the EWS gene translocation with a dual color, break apart probe (Vysis company). RT-PCR was used to detect t (11; 22) (q24; q12) and t (21; 22) (q22; q12) fusion transcripts.


RESULTS
Among 10 cases of ES/PNET, the EWS-FLI1 fusion transcript was detected in 8 by RT-PCR. EWS-ERG fusion transcript was not detected in any of the cases. EWS gene translocation was found in 9 of 10 cases by FISH.


CONCLUSIONS
Interphase FISH and RT-PCR can be reliably applied to paraffin-embedded tissues for molecular diagnosis of ES/PNET. Between the two approaches, interphase FISH provides a more sensitive and stable result.
Yang
Yu
Y
Department of Pathology, Health Science Center, Peking University, Beijing 100083, China.
Wang
Hua
H
Wei
Yan-yu
YY
Zhang
Yan
Y
Ma
Xiao-long
XL
Yang
Xiu-ping
XP
Zheng
Jie
J
D000293:Adolescent; D000328:Adult; D002648:Child; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D018241:Neuroectodermal Tumors, Primitive, Peripheral / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012512:Sarcoma, Ewing / Q000175:diagnosis* / Q000235:genetics