Bockmühl U, Petersen I, Schwendel A, Dietel M
Hals-, Nasen-, Ohrenklinik, Virchow-Klinikum, Humboldt-Universität zu Berlin.
Laryngorhinootologie. 1996 Jul;75(7):408-14. doi: 10.1055/s-2007-997605.
Comparative Genomic Hybridization (CGH) is a novel cytogenetic method that allows the comprehensive analysis of a tumor genome for DNA gains and losses.
CGH was performed on genomic DNA extracted from 14 primary head and neck squamous cell carcinomas. Equal amounts of biotin-labeled tumor DNA and digoxigenin-labeled normal reference DNA were hybridized to normal metaphase chromosomes. The tumor DNA was visualized with fluorescein (FITC) and the normal DNA with rhodamine (TRITC) and detected in a fluorescence microscope. The signal intensities of the different fluorochromes were quantitated as gray levels along the single chromosomes. The over-and underrepresented DNA segments were quantified by computation of FITC/TRITC ratio images and average ratio profiles.
Consensus deletion regions were most frequently observed on chromosome arms 3p (14 cases), 9p (11), 13q (10), 18q (10), 5q (9), 4q (9), 4p (7), 11q (7), 6q (6), 8p (6), and 11p (6). Copy number increases were identified for chromosomes 3q (11), 16p (9), 17q (9), 19p (8), 19q (7), 22 (7), 1p (6), 8q (6), 9q (6), and 20q (6). Particularly, the 3q isochromosome formation (3p loss/3q amplification), found in 10 cases, was a basic alteration. In addition, 11 tumors showed a 11 q13 amplification.
CGH analysis allows the identification of recurrent genetic alterations in head and neck squamous cell carcinomas that will be associated with specific tumor phenotypes like metastatic behavior and prognosis.
比较基因组杂交(CGH)是一种新型细胞遗传学方法,可对肿瘤基因组的DNA增减情况进行全面分析。
对从14例原发性头颈部鳞状细胞癌中提取的基因组DNA进行CGH分析。将等量的生物素标记的肿瘤DNA和地高辛配基标记的正常参照DNA与正常中期染色体杂交。用荧光素(FITC)使肿瘤DNA可视化,用罗丹明(TRITC)使正常DNA可视化,并在荧光显微镜下进行检测。沿单条染色体将不同荧光染料的信号强度定量为灰度级。通过计算FITC/TRITC比率图像和平均比率图谱对DNA片段的增减进行定量。
在3p(14例)、9p(11例)、13q(10例)、18q(10例)、5q(9例)、4q(9例)、4p(7例)、11q(7例)、6q(6例)、8p(6例)和11p(6例)染色体臂上最常观察到共有缺失区域。在3q(11例)、16p(9例)、17q(9例)、19p(8例)、19q(7例)、22(7例)、1p(6例)、8q(6例)、9q(6例)和20q(6例)染色体上发现拷贝数增加。特别地,在10例病例中发现的3q等臂染色体形成(3p缺失/3q扩增)是一种基本改变。此外,11例肿瘤显示11q13扩增。
CGH分析可识别头颈部鳞状细胞癌中反复出现的基因改变,这些改变将与转移行为和预后等特定肿瘤表型相关。