Antonescu Cristina R, Wu Kai, Xing Guoliang Leon, Cao Manqiu, Turpaz Yaron, Leversha Margaret A, Hubbell Earl, Maki Robert G, Miyada C Garrett, Pillai Raji
Department of Pathology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Cancer Inform. 2008;6:59-75. doi: 10.4137/cin.s387. Epub 2008 Mar 27.
We report a method, Expression-Microarray Copy Number Analysis (ECNA) for the detection of copy number changes using Affymetrix Human Genome U133 Plus 2.0 arrays, starting with as little as 5 ng input genomic DNA. An analytical approach was developed using DNA isolated from cell lines containing various X-chromosome numbers, and validated with DNA from cell lines with defined deletions and amplifications in other chromosomal locations. We applied this method to examine the copy number changes in DNA from 5 frozen gastrointestinal stromal tumors (GIST). We detected known copy number aberrations consistent with previously published results using conventional or BAC-array CGH, as well as novel changes in GIST tumors. These changes were concordant with results from Affymetrix 100K human SNP mapping arrays. Gene expression data for these GIST samples had previously been generated on U133A arrays, allowing us to explore correlations between chromosomal copy number and RNA expression levels. One of the novel aberrations identified in the GIST samples, a previously unreported gain on 1q21.1 containing the PEX11B gene, was confirmed in this study by FISH and was also shown to have significant differences in expression pattern when compared to a control sample. In summary, we have demonstrated the use of gene expression microarrays for the detection of genomic copy number aberrations in tumor samples. This method may be used to study copy number changes in other species for which RNA expression arrays are available, e.g. other mammals, plants, etc., and for which SNPs have not yet been mapped.
我们报告了一种名为表达微阵列拷贝数分析(ECNA)的方法,该方法使用Affymetrix人类基因组U133 Plus 2.0阵列,从低至5 ng的输入基因组DNA开始检测拷贝数变化。我们开发了一种分析方法,使用从含有不同X染色体数目的细胞系中分离的DNA,并通过来自在其他染色体位置具有明确缺失和扩增的细胞系的DNA进行验证。我们应用此方法检查了5例冷冻胃肠道间质瘤(GIST)DNA中的拷贝数变化。我们检测到了与先前使用传统或BAC阵列比较基因组杂交(CGH)发表的结果一致的已知拷贝数畸变,以及GIST肿瘤中的新变化。这些变化与Affymetrix 100K人类单核苷酸多态性(SNP)定位阵列的结果一致。这些GIST样本的基因表达数据先前已在U133A阵列上生成,这使我们能够探索染色体拷贝数与RNA表达水平之间的相关性。在GIST样本中鉴定出的一种新畸变,即1q21.1上包含PEX11B基因的先前未报道的增益,在本研究中通过荧光原位杂交(FISH)得到证实,并且与对照样本相比,其表达模式也显示出显著差异。总之,我们证明了使用基因表达微阵列检测肿瘤样本中的基因组拷贝数畸变。该方法可用于研究其他有RNA表达阵列的物种(例如其他哺乳动物、植物等)以及尚未定位SNP的物种中的拷贝数变化。