MOLECULAR CYTOGENETICS OF GENE AMPLIFICATION

利用染色体显微切割，cDNA微阵列分析和CGH结果产生的DNA探针，我们已经确定了一系列涉及人类乳腺癌，卵巢癌和前列腺癌以及其他良性和恶性疾病的染色体区域。为了从这些区域分离候选靶基因，我们开发了基于染色体显微切割和整合基因组作图的技术，包括DNA微阵列分析。这些技术正被用于从乳腺癌、卵巢癌、肉瘤和其他疾病的扩增区域中分离候选基因。先前描述的方法说明了开发快速技术用于鉴定在一系列关键人类肿瘤中扩增的基因的重要性。除了识别已知的基因扩增位点外，我们还鉴定了几个以前未鉴定的基因。我们目前专注于乳腺癌，目前的技术正在为肿瘤进展期间发生的染色体畸变提供新的见解。候选染色体区域和其中包含的基因正在被确定。候选基因的作用机制将通过基于基因转移、基因失活和生物化学技术的模型系统进行研究。此外，我们正在开发一个全面的数据库扩增的染色体区域在软组织和骨肉瘤使用微阵列基于CGH。随着这项技术的成熟，我们已经开始探索其应用于检测癌症中的丢失区域和识别组成染色体异常。这是一个强有力的方法，用于识别在疾病发生或进展中起因果作用的基因，并可能提供新的预后标志物以及治疗靶点。

Using DNA probes generated by chromosome microdissection, cDNA microarray analysis, and CGH findings, we have identified a series of chromosomal regions involved in human breast, ovarian and prostate cancer as well as other benign and malignant diseases. In order to isolate candidate target genes from these regions, we have developed technologies based on chromosome microdissection, and integrated genome mapping, including DNA microarray analysis. These technologies are being utilized to isolate candidate genes from amplified regions in breast cancer, ovarian cancer, sarcomas and other diseases. The methodology previously described illustrates the importance of developing rapid techniques for the identification of genes amplified in a series of key human tumors. In addition to recognizing known sites of gene amplification, we have identified several previously unidentified genes. We are currently focusing on breast cancer where current technologies are providing new insights into the chromosome aberrations which occur during tumor progression. Candidate chromosome regions and genes contained within them are being identified. The mechanism of action of candidate genes will be studied with model systems based on gene transfer, gene inactivation, and biochemical techniques. In addition, we are developing a comprehensive database of amplified chromosomal regions in soft tissue and bone sarcomas using microarray based CGH. As this technology has matured, we have begun to explore its application to the detection of regions of loss in cancer and to the identification of constitutional chromosome anomalies. This is a powerful approach for the identification of genes playing a causal role in disease genesis or progression and may provide novel prognostic markers as well as therapeutic targets.