Williams G H, Rooney S, Thomas G A, Cummins G, Williams E D
Department of Histopathology, University of Cambridge, Addenbrookes Hospital, UK.
Br J Cancer. 1996 Aug;74(4):585-9. doi: 10.1038/bjc.1996.405.
Activation of the RET tyrosine kinase domain occurs in a proportion of thyroid papillary carcinomas. Three chromosomal rearrangements have been described, of which PTC1 is the commonest. Wide differences (2.5-25%) in frequency of PTC1 in different populations have been reported; it is not clear whether these are due to environmental factors, racial differences or technical reasons. We have developed a simple and rapid reverse transcriptase nested polymerase chain reaction (RT-nPCR) method enabling the detection of gene expression from single 5 microns sections of formalin-fixed paraffin wax-embedded archival material. We have applied this approach to detect expression of the RET tyrosine kinase domain, allowing identification of RET activation resulting from any rearrangement, whether characterised or not, or from overexpression. A retrospective study was performed on 22 adult and 21 childhood papillary carcinomas. Thirteen of 22 (59%) adult and 10 of 21 (48%) childhood carcinomas showed evidence of RET activation, demonstrating a major role for the RET oncogene in UK thyroid papillary carcinogenesis. This study also shows a similar frequency of RET activation in both children and adults. The use of a technique that allows reliable amplification of RNA from archival material, using primers chosen in different exons so that amplified products are readily distinguished from genomic DNA, will allow correlation of translocations and chromosomal rearrangements with a variety of specific tumour types.
RET 酪氨酸激酶结构域的激活在一部分甲状腺乳头状癌中出现。已描述了三种染色体重排，其中 PTC1 最为常见。不同人群中 PTC1 的频率差异很大（2.5% - 25%）；尚不清楚这些差异是由于环境因素、种族差异还是技术原因。我们开发了一种简单快速的逆转录巢式聚合酶链反应（RT - nPCR）方法，能够从福尔马林固定石蜡包埋存档材料的单个 5 微米切片中检测基因表达。我们应用这种方法来检测 RET 酪氨酸激酶结构域的表达，从而能够识别由任何重排（无论是否已明确其特征）或过表达导致的 RET 激活。对 22 例成人和 21 例儿童乳头状癌进行了回顾性研究。22 例成人癌中的 13 例（59%）和 21 例儿童癌中的 10 例（48%）显示出 RET 激活的证据，表明 RET 癌基因在英国甲状腺乳头状癌发生中起主要作用。这项研究还表明儿童和成人中 RET 激活的频率相似。使用一种能够从存档材料中可靠扩增 RNA 的技术，该技术使用在不同外显子中选择的引物，以便扩增产物能轻易与基因组 DNA 区分开来，这将使易位和染色体重排与各种特定肿瘤类型相关联。
