Speicher M R, Jauch A, Walt H, du Manoir S, Ried T, Jochum W, Sulser T, Cremer T
Department of Human Genetics, University of Heidelberg, Germany.
Am J Pathol. 1995 Jun;146(6):1332-40.
We present a strategy for the evaluation of numerical copy number changes of DNA segments within a solid tumor genome that allows the correlation of microscopic phenotype with genotype in formalin-fixed, paraffin-embedded tumor material. Cells from a human testicular germ cell tumor and adjacent tissue areas with normal seminiferous tubules were selected separately from microscopically analyzed histological tissue sections, and DNA was extracted from the selected areas. After universal DNA amplification, the amplification products were subjected to comparative genomic hybridization. The results confirmed balanced chromosome copy numbers for the normal tissue area, although the analysis of the tumor tissue area revealed numerous gains and losses of chromosome segments. The comparative genomic hybridization results were used to select DNA probes for interphase cytogenetics on serial sections. We conclude that this technique allows the screening of selected tissue areas for numerical DNA alterations, thus enabling a direct phenotype-genotype comparison.
我们提出了一种评估实体瘤基因组内DNA片段数字拷贝数变化的策略，该策略可使福尔马林固定、石蜡包埋的肿瘤材料中的微观表型与基因型相关联。从经显微镜分析的组织学组织切片中分别选取来自人类睾丸生殖细胞肿瘤的细胞和具有正常生精小管的相邻组织区域，并从所选区域提取DNA。进行通用DNA扩增后，将扩增产物进行比较基因组杂交。结果证实正常组织区域的染色体拷贝数平衡，尽管对肿瘤组织区域的分析显示出许多染色体片段的增加和缺失。比较基因组杂交结果用于在连续切片上选择用于间期细胞遗传学的DNA探针。我们得出结论，该技术可筛选选定组织区域的DNA数字改变，从而实现直接的表型-基因型比较。
