ERBB2 AMPLIFICATION IN BREAST-CANCER ANALYZED BY FLUORESCENCE INSITU HYBRIDIZATION
ERBB2 AMPLIFICATION IN BREAST-CANCER ANALYZED BY FLUORESCENCE INSITU HYBRIDIZATION
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DOI：
10.1073/pnas.89.12.5321
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发表时间：
1992-06-15
影响因子：
11.1
通讯作者：
GRAY, JW
中科院分区：
文献类型：
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作者：
KALLIONIEMI, OP;KALLIONIEMI, A;GRAY, JW
我们使用荧光原位杂交（FISH）分析ERBB2癌基因拷贝数、扩增水平（这里定义为ERBB2拷贝数与第17号染色体着丝粒拷贝数之比），以及扩增基因在乳腺癌细胞系和未培养的原发性乳腺癌中的分布。在10种乳腺癌细胞系中，FISH测定的ERBB2相对拷贝数与Southern blot结果高度相关（r = 0.98），当两种方法使用同一参考位点的探针时。细胞系的中期分析表明，扩增的ERBB2拷贝总是发生在染色体内集群中，但这些集群的数量和染色体位置在细胞系之间有所不同。在显示ERBB2扩增的原发肿瘤的间期核中（10/44），ERBB2拷贝被视为一到四个簇，也提示染色体内定位。无论平均扩增水平如何，所有这些肿瘤都含有高度扩增的细胞亚群，每个细胞至少有25个，有时超过100个ERBB2拷贝。未显示FISH扩增的肿瘤（34/44）平均有1至5个ERBB2拷贝随机分布在细胞核中，完全缺乏高拷贝水平的细胞。原发肿瘤的FISH结果与槽印迹扩增结果和免疫组化过表达检测结果一致。通过FISH对ERBB2扩增进行定量分析，可以改善基于扩增模式和检测大量扩增肿瘤细胞亚群的预后评估。
We illustrate the use of fluorescence in situ hybridization (FISH) for analysis of ERBB2 oncogene copy number, the level of amplification (here defined as the ratio of ERBB2 copy number to copy number of chromosome 17 centromeres), and the distribution of amplified genes in breast cancer cell lines and uncultured primary breast carcinomas. The relative ERBB2 copy number determined by FISH in 10 breast cancer cell lines correlated strongly with Southern blot results (r = 0.98) when probes for an identical reference locus were used in the two methods. Metaphase analysis of cell lines showed that amplified ERBB2 copies always occurred in intrachromosomal clusters but that the number and chromosomal location of these clusters varied among the cell lines. In interphase nuclei of primary tumors showing ERBB2 amplification (10/44), ERBB2 copies were seen as one to four clusters, also suggesting intrachromosomal localization. Regardless of the average level of amplification, all these tumors contained highly amplified cell subpopulations with at least 25, and sometimes more than 100, ERBB2 copies per cell. Tumors that did not show amplification by FISH (34/44) had an average of one to five ERBB2 copies scattered randomly in the nuclei and completely lacked cells with high copy levels. FISH results on primary tumors were concordant with slot blot results on amplification and with immunohistochemical detection of over-expression. Quantitative analysis of ERBB2 amplification by FISH may improve prognostic assessments based on the pattern of amplification and detection of heavily amplified tumor cell subpopulations.