Lahr G, Stich M, Schütze K, Blümel P, Pösl H, Nathrath W B
Laser Laboratory and Molecular Biology, First Medical Department, Municipal and Teaching Hospital München-Harlaching, Munich, Germany.
Pathobiology. 2000;68(4-5):218-26. doi: 10.1159/000055927.
The purpose of this study was to investigate the value of the expression of the RET oncogene (rearranged during transfection) in papillary thyroid carcinomas (PTC) and its variants in the differential diagnosis of thyroid neoplasias. According to the literature RET oncogene activation by chromosomal rearrangements has been exclusively implicated in PTCs.
To establish the incidence of RET activation in PTCs we used 5- to 10-microm sections from archival paraffin blocks. Either parts of the tissue slices were manually dissected or a few distinct cells were microdissected by laser-mediated manipulation with the Robot-MicroBeam system. RNA was extracted from paraffin-embedded thyroid tumors and the corresponding normal tissue. RT and nested PCR were performed using primers for RET/PTC1, PTC2 and PTC3, or for RET exons 12 and 13. PCR products were resolved by gel electrophoresis.
We detected RET transcription in approximately 85% of the PTCs including follicular variants and in isolated cells of the same tissues, but not in nonmalignant thyroid tissue.
Our method may serve as an additional diagnostic tool to characterize ambiguous neoplasias and to identify especially nonpapillary, i.e. follicular tumors, as papillary carcinomas. Additionally, this study has demonstrated that expressed genes can be analyzed from routine histopathological tissue slides or pooled single cells. Large retrospective studies can also be performed with this method.
本研究旨在探讨转染期间重排(RET)癌基因在甲状腺乳头状癌(PTC)及其变体中表达的价值,以及其在甲状腺肿瘤鉴别诊断中的作用。根据文献,染色体重排激活RET癌基因仅与PTC有关。
为确定PTC中RET激活的发生率,我们使用了存档石蜡块中5至10微米厚的切片。组织切片的部分区域通过手动解剖,或使用机器人微束系统通过激光介导操作对少数离散细胞进行显微切割。从石蜡包埋的甲状腺肿瘤及其相应的正常组织中提取RNA。使用针对RET/PTC1、PTC2和PTC3或RET外显子12和13的引物进行逆转录(RT)和巢式聚合酶链反应(PCR)。PCR产物通过凝胶电泳进行分离。
我们在大约85%的PTC(包括滤泡变体)及其相同组织的单个细胞中检测到RET转录,但在非恶性甲状腺组织中未检测到。
我们的方法可作为一种辅助诊断工具,用于鉴别不明确的肿瘤,并将特别是非乳头状(即滤泡性)肿瘤识别为乳头状癌。此外,本研究表明,可以从常规组织病理学组织切片或汇集的单个细胞中分析表达的基因。也可以使用这种方法进行大型回顾性研究。