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首页> 外文期刊>Pathobiology: journal of immunopathology, molecular and cellular biology >Detection of gene amplification in intraductal and infiltrating breast cancer by laser-assisted microdissection and quantitative real-time PCR.
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Detection of gene amplification in intraductal and infiltrating breast cancer by laser-assisted microdissection and quantitative real-time PCR.

机译：激光辅助显微解剖和定量实时PCR检测导管内和浸润性乳腺癌中的基因扩增。

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摘要

Gene amplification is one essential mechanism leading to oncogene activation which is supposed to play a major role in the pathogenesis of invasive breast cancer. However, using standard methodologies the detection of gene amplifications has been limited especially in small-sized lesions, like pre-invasive precursor lesions. The combination of two novel technologies, laser-based microdissection and quantitative real-time PCR, facilitates the detection of low-level amplifications in morphologically defined lesions. As a model system we investigated in situ breast cancer (ductal carcinoma in situ, DCIS) classified according to the morphology-based Van Nuys grading system for amplification of growth-regulatory genes. In this study 83 formalin-fixed, paraffin-embedded archival DCIS specimens were examined after laser-based microdissection by quantitative real-time PCR using the TaqMan detection system for amplification of the c-erbB2, topoisomerase IIalpha, c-myc and cyclinD1 gene. In a subset of 17 DCIS with adjacent infiltrating tumour components we compared intraductal and invasive tumour components in parallel for differences in amplification status. The combination of these new techniques represents an excellent tool to gain new insights into carcinogenesis by analyzing genetic alterations in morphologically identified heterogeneous lesions in breast cancer progression within the very same specimen or even tissue slide. Copyright 2001 S. Karger AG, Basel
机译：基因扩增是导致癌基因激活的一种重要机制，该机制可能在浸润性乳腺癌的发病机理中起主要作用。但是，使用标准方法，基因扩增的检测受到了限制，尤其是在较小的病变中，如浸润前体病变。基于激光的显微解剖和实时定量PCR这两种新技术的结合，有助于在形态学上确定的病变中检测低水平的扩增。作为模型系统，我们研究了原位乳腺癌（导管原位癌，DCIS），该原位乳腺癌是根据基于形态学的Van Nuys分级系统进行分类的，用于扩增生长调节基因。在这项研究中，使用TaqMan检测系统通过定量实时PCR扩增了c-erbB2，拓扑异构酶IIalpha，c-myc和cyclinD1基因，对基于激光的显微解剖后的83份福尔马林固定，石蜡包埋的档案DCIS标本进行了检查。在具有相邻浸润性肿瘤成分的17个DCIS的子集中，我们比较了并行的导管内和浸润性肿瘤成分的扩增状态差异。这些新技术的结合代表了一种极好的工具，可以通过分析在同一样本甚至组织切片中乳腺癌进展过程中形态学确定的异质性病变的遗传改变来获得有关癌变的新见解。版权所有2001 S. Karger AG，巴塞尔

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