Detection of EWSR1 translocation with nuclear extraction-based fluorescence in situ hybridization for diagnosis of Ewing's sarcoma/primitive neuroectodermal tumor.
OBJECTIVE
To investigate nuclear extraction-based fluorescence in situ hybridization (NE-FISH) in EWSR1 translocation of Ewing's sarcoma/primitive neuroectodermal tumor (ES/PNET) in formalin-fixed, paraffin-embedded (FFPE) tissue, compared to thin-section FISH (TS-FISH) and nested reverse-transcription polymerase chain reaction (RT-PCR).


METHODS
Nuclei of the tumor cells and total ribonucleic acid (RNA) were extracted from 10 cases of ES/PNET. NE-FISH and TS-FISH were used to analyze the EWS gene translocation. RT-PCR was used to detect t(11;22)(q24;q12) and t(21;22)(q22;q12) fusion transcripts.


RESULTS
Of 10 cases, 9 (90%) were abnormal by NE-FISH and TS-FISH. The average rate of abnormal split signals detected by NE-FISH was significantly higher than by TS-FISH. We analyzed 10 ES/PNET cases and 8 various normal human tissues by NE-FISH and nested RT-PCR. Fusion signals were found in 9 (90%) ES/PNET cases by NE-FISH and 8 (80%) by nested RT-PCR. Of 8 normal tissues, none was abnormal by either FISH or RT-PCR.


CONCLUSIONS
NE-FISH is more reliable than TS-FISH in detecting EWSR1 translocation of ES/PNET in FFPE tissue, although the FISH techniques were concordant. Nested RT-PCR could be more sensitive than conventional RT-PCR. A combination of NE-FISH and nested RT-PCR would improve accuracy of molecular diagnosis of ES/PNET.
Yang
Yu
Y
Department of Pathology, Health Science Center, Peking University, Beijing, China.
Zhang
Lanjing
L
Wei
Yanyu
Y
Wang
Hua
H
Xiong
Wei
W
Chen
Zheng
Z
Hes
Ondrej
O
Zheng
Jie
J
D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D001859:Bone Neoplasms / Q000175:diagnosis* / Q000235:genetics; D002148:Calmodulin-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004273:DNA, Neoplasm / Q000032:analysis; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D018241:Neuroectodermal Tumors, Primitive, Peripheral / Q000175:diagnosis* / Q000235:genetics; D034802:RNA-Binding Protein EWS; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012512:Sarcoma, Ewing / Q000175:diagnosis* / Q000235:genetics; D014178:Translocation, Genetic
D014408:Biomarkers, Tumor; D002148:Calmodulin-Binding Proteins; D004273:DNA, Neoplasm; C500986:EWSR1 protein, human; D034802:RNA-Binding Protein EWS; D016601:RNA-Binding Proteins