Intratumoral heterogeneity in breast carcinoma revealed by laser-microdissection and comparative genomic hybridization
Intratumoral heterogeneity in breast carcinoma revealed by laser-microdissection and comparative genomic hybridization
复制标题
DOI：
10.1016/s0165-4608(98)00205-2
复制
发表时间：
1999-04-15
影响因子：
--
通讯作者：
Werner, M
中科院分区：
文献类型：
--
作者：
Aubele, M;Mattis, A;Werner, M
为了评估乳腺癌中潜在的细胞遗传学异质性，在石蜡切片中研究了几个小细胞群（每个由20至50个细胞组成）。BE激光显微切割，每例取3~7个细胞群。用简并寡核苷酸引物PCR（DOP-PCR）扩增DNA，并用CGH分析染色体的获得和丢失。本文报告两例导管浸润性乳腺癌，其中一例有两个淋巴结转移。为了比较小样本的结果，还对从三至五个连续切片（10（7）至10（6）个细胞）的肿瘤区域分离的DNA进行了CGH。在显微切割的肿瘤样本中观察到的畸变是多个的，涉及多达14个不同的染色体或亚染色体区域。最常见的变化是染色体滞后（14/20）和20 q（16/20）的增加和13 q（12/20）的丢失。一些畸变很少被检测到（例如，2p上的损失，8q上的增益）。比较原发肿瘤和淋巴结转移瘤的染色体失衡，原发肿瘤和相应的转移瘤之间的染色体失衡比原发肿瘤和淋巴结转移瘤之间的染色体失衡更一致。激光显微切割的样品一般表现出更多的染色体畸变比DNA分离的几个肿瘤切片。我们的CGH结果证实了荧光原位杂交（FISH）的染色体区域的着丝粒1和20，和20q13。此外，31个样本的微卫星分析证实了我们对选定的染色体区域2p和11q的CGH结果。如果cc-m可以得出结论，在原发性乳腺肿瘤以及相应的淋巴结转移瘤中存在明显的瘤内异质性。显微切割和CGH的结合使我们能够从重要的克隆中检测出细胞遗传学畸变，这些克隆在分析从大量细胞中提取的DNA时被遗漏。(C)Elsevier Science Inc.，1999. All rights reserved.
To evaluate the potential cytogenetic heterogeneity in breast carcinoma, several small cell groups (each consisting of 20 to 50 cells) were investigated within paraffin sections. BE laser-microdissection, three to seven cell groups rt ere taken per case. The DNA was amplified by degenerate oligonucleotide primed PCR (DOP-PCR) and the samples tr ere analyzed by CGH for chromosomal gains and losses. Two ductal invasive breast carcinomas, one of them with two lymphnode metastases, rr ere investigated. To compare the results from the small samples, CGH was also performed on DNA isolated from the tumorous regions of three to five serial sections (10(7) to 10(6) cells). The aberrations observed in the microdissected tumor samples were multiple and involved up to 14 different chromosomal or subchromosomal regions. The most frequent changes were gains on chromosomes lag (14/20) and 20q (16/20), and loss on 13q (12/20). Some aberrations have rarely been detected (e.g., loss on 2p, gain on 8q). Comparing chromosomal imbalances in primary tumors and lymph node metastases, more consistent changes rr ere found between the primary tumor and its corresponding metastases than between both primary tumors. The laser-microdissected samples in general showed more chromosomal aberrations than DNA isolated from several tumor sections. Our CGH results ri ere confirmed by fluorescence in situ hybridization (FISH) for the chromosomal regions of centromere 1 and 20, and 20q13. In addition, microsatellite analyses on 31 samples confirmed our CGH findings for selected chromosome regions 2p and 11q. If cc-m be concluded that there is a distinct intratumoral heterogeneity in primary breast tumors as well as in the corresponding lymph node metastases. The combination of microdissection and CGH enabled us to detect cytogenetic aberrations from important clones which are missed rr hen analyzing DNA extracted from large cell numbers. (C) Elsevier Science Inc., 1999. All rights reserved.