Trent J M, Bittner M, Zhang J, Wiltshire R, Ray M, Su Y, Gracia E, Meltzer P, De Risi J, Penland L, Brown P
Laboratory of Cancer Genetics, National Center for Human Genome Research, National Institutes of Health, Bethesda, MD 20892-4470, USA.
Clin Exp Immunol. 1997 Jan;107 Suppl 1:33-40.
Chromosome abnormalities in human malignancies have identified the genomic location of several important growth-regulatory genes, including cellular oncogenes and tumour suppressor genes. Melanomas are characterized by recurring chromosome alterations, and it is important to identify those genes whose altered expression may be causally related to melanocytic transformation. This short report presents an overview of strategies used which combine the materials and technologies of the Human Genome Project with clinically directed studies of melanoma biology. The Human Genome Project combines various technologies, including cytogenetic, physical mapping, genetic mapping and DNA sequencing, in order to identify all of the human genes, but especially the 4000 estimated to contribute to human disease. This report focuses first on advances in genome technology that provide information on chromosome rearrangements and DNA copy number changes. This includes a discussion of chromosome microdissection as well as the microexcision of tissue specimens to gain insights into chromosome regions altered in association with melanocyte transformation. Next, there is a brief discussion of the generation and characterization of subtracted cDNA sublibraries which allow the identification of genes uniquely expressed in association with the transformed phenotype of human melanoma cells. Finally, we briefly discuss the feasibility of using a recently developed system for parallel examination of multiple genes based upon robotic printing of cDNAs on glass slides, and simultaneous two-colour fluorescence hybridization to study the expression patterns of cDNAs for their association with melanoma tumour suppression. The combination of these varied molecular technologies may provide insights into previously unrecognized genes involved causally in the pathobiology of this important neoplasm, and may provide new targets for clinical intervention.
人类恶性肿瘤中的染色体异常已确定了几个重要生长调节基因的基因组位置，包括细胞癌基因和肿瘤抑制基因。黑色素瘤的特征是反复出现染色体改变，因此确定那些表达改变可能与黑素细胞转化有因果关系的基因非常重要。本简短报告概述了所采用的策略，这些策略将人类基因组计划的材料和技术与黑色素瘤生物学的临床定向研究相结合。人类基因组计划结合了多种技术，包括细胞遗传学、物理图谱绘制、遗传图谱绘制和DNA测序，以识别所有人类基因，尤其是估计有4000个与人类疾病相关的基因。本报告首先关注基因组技术的进展，这些进展提供了有关染色体重排和DNA拷贝数变化的信息。这包括对染色体显微切割以及组织标本微切除的讨论，以便深入了解与黑素细胞转化相关的染色体区域变化。接下来，简要讨论了扣除cDNA亚文库的生成和特征，该文库可用于识别与人类黑色素瘤细胞转化表型相关的独特表达基因。最后，我们简要讨论了使用最近开发的系统基于在载玻片上机器人打印cDNA并行检测多个基因以及同时进行双色荧光杂交来研究cDNA表达模式与黑色素瘤肿瘤抑制相关性的可行性。这些不同分子技术的结合可能会为以前未被认识的、在这种重要肿瘤的病理生物学中起因果作用的基因提供深入了解，并可能为临床干预提供新的靶点。
