REVERSE-TRANSCRIPTASE PCR AMPLIFICATION OF EWS/FLI-1 FUSION TRANSCRIPTS AS A DIAGNOSTIC-TEST FOR PERIPHERAL PRIMITIVE NEUROECTODERMAL TUMORS OF CHILDHOOD
REVERSE-TRANSCRIPTASE PCR AMPLIFICATION OF EWS/FLI-1 FUSION TRANSCRIPTS AS A DIAGNOSTIC-TEST FOR PERIPHERAL PRIMITIVE NEUROECTODERMAL TUMORS OF CHILDHOOD
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DOI：
10.1097/00019606-199300020-00024
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发表时间：
1993-09-01
影响因子：
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通讯作者：
TRICHE, TJ
中科院分区：
文献类型：
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作者：
SORENSEN, PHB;LIU, XF;TRICHE, TJ
儿童周围原始神经外胚层肿瘤 (pPNET)，包括尤文肉瘤、周围神经上皮瘤和阿斯金肿瘤，通常给解剖病理学家带来重大的诊断挑战。这些肿瘤的一个一致特征是肿瘤细胞中存在 t(11;22)(q24;q12)，这种易位可用作这组肿瘤的标记物。最近对t(11;22)断点的克隆揭示了染色体11q24上的人类FLI-1基因与22q12上一个名为EWS的未知功能基因的融合，并且已检测到融合转录本。这些发现提出了使用分子遗传学分析作为诊断 pPNET 的工具的可能性。为此，我们使用 EWS 和 FLI-1 特异性引物通过逆转录酶聚合酶链式反应 (RT-PCR) 测试了 pPNET 是否存在 EWS/FLI-1 融合转录本。使用此技术，10 个 pPNET 细胞系中的 8 个（80%）对扩增产物呈阳性。这些结果得到了 Southern 分析的证实，Southern 分析揭示了使用基因组 EWS 探针在所有八种阳性细胞系中进行的 EWS 重排。然后，我们测试了 20 个原发性 pPNET 肿瘤，并通过 RT-PCR 在其中 18 个病例 (90%) 中鉴定了融合转录本。 PCR 产物的克隆和测序证实了这些产物中存在 EWS 和 FLI-1 序列。此外，该技术在一系列非 pPNET 儿科实体瘤中未检测到融合转录本。因此，通过 RT-PCR 检测 EWS/FLI-1 融合转录本为 pPNET 的诊断提供了一种新的辅助工具。
The peripheral primitive neuroectodermal tumors (pPNETs) of childhood, including Ewing's sarcoma, peripheral neuroepithelioma, and Askin's tumor, often present significant diagnostic challenges for the anatomic pathologist. One consistent feature of these tumors is the presence of the t(11;22)(q24;q12) in tumor cells, and this translocation has been useful as a marker for this group of tumors. The recent cloning of the t(11;22) breakpoint has revealed the fusion of the human FLI-1 gene on chromosome 11q24 with a gene of unknown function called EWS on 22q12, and fusion transcripts have been detected. These findings have raised the possibility of using molecular genetic analysis as a tool to diagnose pPNETs. To this end, we have tested pPNETs for the presence of EWS/FLI-1 fusion transcripts by reverse transcriptase-polymerase chain reaction (RT-PCR) using EWS and FLI-1 specific primers. Eight (80%) of 10 pPNET cell lines were positive for amplified products using this technique. These results were confirmed by Southern analysis, which revealed rearrangements of EWS using genomic EWS probes in all eight positive cell lines. We then tested 20 primary pPNET tumors, and identified fusion transcripts by RT-PCR in 18 (90%) of these cases. Cloning and sequencing of PCR products confirmed the presence of EWS and FLI-1 sequences in these products. Furthermore, fusion transcripts were not detected by this technique in a series of non-pPNET pediatric solid tumors. Detection of EWS/FLI-1 fusion transcripts by RT-PCR therefore provides a novel adjunctive tool in the diagnosis of pPNETs.