Balazs M, Matsumura K, Moore D, Pinkel D, Gray J W, Waldman F M
Department of Laboratory Medicine, University of California San Francisco 94143-0808, USA.
Cytometry. 1995 May 1;20(1):62-73. doi: 10.1002/cyto.990200110.
We have used fluorescence in situ hybridization (FISH) with chromosome-specific probes and immunofluorescent detection of in vivo bromodeoxyuridine (BrdUrd) incorporation to evaluate simultaneously numerical chromosome aberrations and proliferative activity of breast cancers. The number of distinct hybridization domains specific for repetitive pericentromeric sequences on chromosomes 1, 7, 11, 15, 17, and X was used as an indicator of copy number of these chromosomes in interphase tumor cells from 23 human breast cancers. Every tumor analyzed showed a heterogeneous distribution of copy number for at least one chromosome type. The copy number distribution for different chromosomes within a tumor frequently showed differing patterns. Major cell populations showing monosomy were relatively rare, occurring only in five cases for chromosome 17, once for chromosome 1, and once for chromosome 15. Flow cytometric analysis of DNA ploidy correlated well with FISH analysis, although flow cytometry failed to detect aneuploidy when only a few chromosomes were affected. To determine whether cell populations with different chromosomal copy numbers have identical proliferation characteristics in vivo, BrdUrd incorporation and centromeric copy number were detected simultaneously. Comparison of the chromosome copy number distribution in BrdUrd-positive cells vs. the distribution of the entire cell population showed different distributions in seven of the 20 cases analyzed. This study demonstrates the common occurrence of chromosome copy number heterogeneity and suggests that a cell phenotype (proliferation) may be associated with genotypic subpopulations.
我们运用了针对特定染色体的探针进行荧光原位杂交（FISH），并通过免疫荧光检测体内溴脱氧尿苷（BrdUrd）的掺入情况，以同时评估乳腺癌的染色体数目畸变和增殖活性。对于来自23例人类乳腺癌的间期肿瘤细胞，将1号、7号、11号、15号、17号染色体以及X染色体上重复的着丝粒周围序列所特有的不同杂交区域的数量用作这些染色体拷贝数的指标。所分析的每例肿瘤至少有一种染色体类型呈现出拷贝数的异质性分布。肿瘤内不同染色体的拷贝数分布常常呈现出不同的模式。显示单体性的主要细胞群体相对较少，仅在17号染色体的5例、1号染色体的1例以及15号染色体的1例中出现。DNA倍体的流式细胞术分析与FISH分析相关性良好，尽管当只有少数染色体受到影响时，流式细胞术未能检测到非整倍体。为了确定具有不同染色体拷贝数的细胞群体在体内是否具有相同的增殖特征，同时检测了BrdUrd掺入情况和着丝粒拷贝数。在分析的20例病例中，有7例BrdUrd阳性细胞中的染色体拷贝数分布与整个细胞群体的分布不同。这项研究证明了染色体拷贝数异质性的普遍存在，并表明细胞表型（增殖）可能与基因型亚群相关。
