摘要
目的探讨运用荧光原位杂交(FISH)与逆转录聚合酶链反应(RT-PCR)检测尤文肉瘤/原始神经外胚层肿瘤(ES/PNET)石蜡包埋组织特异性染色体易位的临床应用价值。方法收集ES/PNET10例,分离石蜡包埋组织中的肿瘤细胞,利用Vysis公司的EWSR1双色易位分离探针,间期核FISH检测EWS基因的易位。运用RT-PCR检测t(11;22)(q24;q12)和t(21;22)(q22;q12)形成的融合转录子EWS-FLI1和EWS-ERG。结果10例ES/PNET中FISH观察9例有EWS基因的易位。RT-PCR8例检测出EWS-FLI1的表达,没有检测出EWS-ERG的表达。结论利用FISH与RT-PCR检测ES/PNET石蜡包埋组织特异性染色体易位可作为可靠的分子诊断指标;两者相比,FISH敏感性和稳定性要优于RT-PCR。
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. Condusions Interphase FISH and RT-PCR can be reliably applied to paraffin-embedded tissues for molecular diagnosis of ES/PNET. Between the two approaches, interohase FISH provides a more sensitive and stable result.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2006年第6期328-332,共5页
Chinese Journal of Pathology
基金
国家重点基础研究发展规划项目(973)(2002CB513100)
关键词
神经外胚瘤
原始
外周
原位杂交
荧光
逆转录聚合酶链反应
Neuroectodermal tumors, primitive, peripheral
In situ hybridization, fluorescence
Reverse transcription polymerase chain reaction