High-resolution array comparative genomic hybridization of single micrometastatic tumor cells.
High-resolution array comparative genomic hybridization of single micrometastatic tumor cells.
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DOI：
10.1093/nar/gkn101
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发表时间：
2008-04
影响因子：
14.9
通讯作者：
Klein CA
中科院分区：
文献类型：
--
作者：
Fuhrmann C;Schmidt-Kittler O;Stoecklein NH;Petat-Dutter K;Vay C;Bockler K;Reinhardt R;Ragg T;Klein CA
只有少数选定的癌细胞驱动肿瘤进展，并负责治疗抗性。然而，它们的特定基因组特征在很大程度上是未知的，因为高分辨率基因组分析目前仅限于从许多细胞中汇集的DNA。在这里，我们描述了一个协议阵列比较基因组杂交（阵列CGH），它能够检测单细胞中的DNA拷贝数的变化。将基于PCR的全基因组扩增方法与高度纯化的BAC克隆阵列相结合，我们可以准确地确定已知的染色体变化，例如单个白细胞中的21三体以及单细胞系细胞的复杂基因组失衡。在单个T47 D细胞中，当与来自106个细胞的非扩增DNA相比时，在大多数情况下鉴定出小至1-2 Mb的异常区域。最重要的是，在从乳腺癌患者骨髓中分离的单个微转移癌细胞中，我们检索并确认了小至4.4和5 Mb的扩增。因此，单转移性前体细胞的高分辨率基因组分析现在是可能的，并可用于鉴定新的治疗靶基因。
Only few selected cancer cells drive tumor progression and are responsible for therapy resistance. Their specific genomic characteristics, however, are largely unknown because high-resolution genome analysis is currently limited to DNA pooled from many cells. Here, we describe a protocol for array comparative genomic hybridization (array CGH), which enables the detection of DNA copy number changes in single cells. Combining a PCR-based whole genome amplification method with arrays of highly purified BAC clones we could accurately determine known chromosomal changes such as trisomy 21 in single leukocytes as well as complex genomic imbalances of single cell line cells. In single T47D cells aberrant regions as small as 1–2 Mb were identified in most cases when compared to non-amplified DNA from 106 cells. Most importantly, in single micrometastatic cancer cells isolated from bone marrow of breast cancer patients, we retrieved and confirmed amplifications as small as 4.4 and 5 Mb. Thus, high-resolution genome analysis of single metastatic precursor cells is now possible and may be used for the identification of novel therapy target genes.
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DOI：
10.1186/bcr1370
发表时间：
2006
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Shadeo A;Lam WL
通讯作者：
Lam WL
影响因子：
168.9
作者：
Klein, CA;Blankenstein, TJF;Riethmüller, G
通讯作者：
Riethmüller, G
影响因子：
50.3
作者：
Schardt, JA;Meyer, M;Klein, CA
通讯作者：
Klein, CA
影响因子：
4.4
作者：
Frengen, E;Weichenhan, D;de Jong, PJ
通讯作者：
de Jong, PJ
影响因子：
64.8
作者：
Lengauer, C;Kinzler, KW;Vogelstein, B
通讯作者：
Vogelstein, B