Application of fluorescence in-situ hybridization and reverse transcription-polymerase chain reaction in molecular diagnosis of Ewing's sarcoma and primitive neuroectodermal tumor.
- Author:
Yu YANG
1
;
Hua WANG
;
Yan-yu WEI
;
Yan ZHANG
;
Xiao-long MA
;
Xiu-ping YANG
;
Jie ZHENG
Author Information
- Publication Type:Journal Article
- MeSH: Adolescent; Adult; Child; Humans; In Situ Hybridization, Fluorescence; methods; Neuroectodermal Tumors, Primitive, Peripheral; diagnosis; genetics; Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; methods; Sarcoma, Ewing; diagnosis; genetics
- From: Chinese Journal of Pathology 2006;35(6):328-332
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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).
METHODSNuclei 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.
RESULTSAmong 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.
CONCLUSIONSInterphase 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.
