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題名 | Detection of PAX3-FKHR and PAX7-FKHR Fusion Transcripts in Rhabdomyosarcoma by Reverse Transcriptase-Polymerase Chain Reaction Using Paraffin-Embedded Tissue=以反轉錄聚合酶鏈反應方法偵測橫紋肌肉瘤臘埋組織中PAX3-FKHR PAX7-FKHR的交移訊息 |
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作者 | 陳碧芳; 陳孟良; 梁德城; 黃玉文; 劉希哲; 陳淑惠; | 書刊名 | 中華醫學雜誌 |
卷期 | 62:2 1999.02[民88.02] |
頁次 | 頁86-91 |
分類號 | 415.138 |
關鍵詞 | 小泡型橫紋肌肉瘤; FKHR基因; 福馬林固定之臘埋組織; PAX3基因; PAX7基因; 反轉錄聚合酶鏈反應法; Alveolar rhabdomyosarcoma; FKHR gene; Formalin-fixed paraffin-embedded tissue; PAX3 gene; PAX7 gene; Reverse transcriptase-polymerase chain reaction; |
語文 | 英文(English) |
中文摘要 | 背景:小泡型橫紋肌肉瘤具有典型的染色體移位t(2.13)(q35;ql4),移位的基因 位於第 2 對染色體上的 PAX3 和第 13 對染色體上的 FKHR。此外,這類腫瘤極少具有第 1 對染色體 PAX7 和第 13 對染色體 FKHR 基因的移位。有關橫紋肌肉瘤染色體移位之偵測, 大多數實驗室是利用傳統的細胞遺傳學方法、螢光原位雜交法或反轉錄聚合�t鏈反應法,並 且大多需要使用新鮮或冷凍的標本。本實驗利用 17 個福馬林固定後橫紋肌肉瘤的臘埋組織 ,以反轉錄聚合�t鏈反應法來偵測 PAX3-FKHR 和 PAX7-FKHR 的交移訊息。 方法:從福馬林固定後之臘埋組織抽取 RNA, 使用相當於 PAX3、PAXK7 及 FKHR 區域的寡 核��酸引子來偵測 PAX3-FKHR 或 PAX7-FKHR 的交移訊息。此外,聚合�t鏈反應之產物可用 半聚集之方法來增加偵測的敏感度,而再將放大後的片段加以純化,並直接定序以確立其特 異性。 結果:3 個小泡型橫紋肌肉瘤皆可偵測到 PAX3-FKHR 或 PAX7-FKHR 的交移訊息,但其它亞 型,包括 12 個胚胎型或多形性型則無偵測到; 另外,PAX7-FKHR 之交移僅有 1 例胚胎型 橫紋肌肉瘤測得訊息。 結論:以反轉錄聚合醃鏈反應法來偵測橫紋肌肉瘤的 PAXK3-FKHR 交移訊息,也可以應用於 福馬林固定後的臘埋組織,其結果相當準確。當無法得到新鮮或冷凍的組織時,病理科可應 用此法作為輔助性之診斷,以偵測橫紋肌肉瘤染色體之移位。 |
英文摘要 | Background. Alveolar rhabdomyosarcoma (RMS) is associated with a characteristic chromosomal translocation t(2;13)(q35;ql4). The genes involved in this translocation are paired box (PAX)3 on chromosome 2 and forkhead in RMS (FKHR) on chromosome 13. An occasional variant translocation t(l;13)(p36;ql4) affecting PAX7 and FKHR on chromosomes I and 13, respectively, has also been described. Chromosomal translocations in RMS are detected using conventional cytogenetic analysis, fluorescence in situ hybridization (FISH) or reverse transcriptase-polymerase chain reaction (RT-PCR) on fresh or frozen tissue samples. We describe the results of RT-PCR analysis of PAX3-FKHR and PAX7-FKHR chimeric messages in formalin-fixed, paraffin-embedded tissue samples from 17 RMS cases. Methods. RNA was extracted from formalin-fixed, paraffin-embedded RMS tissue. Oligonucleotide primers corresponding to the regions of PAX3, PAX7 and FKHR were used for the detection of PAX3-FKHR and PAX7-FKHR chimeric messages. A seminested PCR of the PCR products was used to increase the sensitivity of detection. The amplified fragments were purified and directly sequenced to confirm the specificity of the methods. Results. The PAX3-FKHR chimeric message was detected in all three cases of alveolar RMS but not in any of the 12 embryonal and two pleomorphic RMS cases. The PAX7-FKHR fusion transcript was detected in one case of embryonal RMS. Conclusions. The results indicate that the RT-PCR assay is a reliable method for the detection of the PAX3-FKHR fusion,transcript of alveolar RMS in formalin-fixed, paraffin- embedded tissue. This simple method enables pathologists to identify chromosomal rearrangements in RMS as a diagnostic aid in cases where fresh or frozen tissue is not available. |
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