Hidden gene amplifications in aggressive B-cell non-Hodgkin lymphomas detected by microarray-based comparative genomic hybridization
Hidden gene amplifications in aggressive B-cell non-Hodgkin lymphomas detected by microarray-based comparative genomic hybridization
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DOI：
10.1038/sj.onc.1206297
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发表时间：
2003-03-06
期刊：
影响因子：
8
通讯作者：
Bentz, M
中科院分区：
文献类型：
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作者：
Wessendorf, S;Schwaenen, C;Bentz, M
DNA扩增是原癌基因激活的重要机制。中期染色体制备的比较基因组杂交（CGH）显示10-20%的B细胞淋巴瘤（B-NHL）扩增。我们分析了一系列的16个侵袭性非霍奇金淋巴瘤的新方法称为矩阵CGH（M-CGH）使用基因组DNA微阵列作为杂交目标。对于M-CGH，构建了一个专用的B细胞淋巴瘤芯片，其中包含496个基因组靶标，涵盖癌基因、肿瘤抑制基因以及B-NHL中经常改变的染色体区域。在16个样品中的10个中，共鉴定出15个DNA扩增。扩增子包括BCL 2、REL、CCND 1、CCND 2、JAK 2、FGF 4和MDM 2。15个扩增中有4个未被染色体CGH检测到。通过荧光原位杂交证实了分别定位于条带2 p13、9 p13-p21和12 q24的扩增子。此外，对于四个基因组扩增的基因，实时定量逆转录聚合酶链反应显示mRNA表达水平升高。这些数据显示了新开发的诊断工具的上级诊断灵敏度。由于只有一小部分的基因组（约1.5%）已被本DNA阵列分析，这是可能的，基因扩增是更常见的侵袭性淋巴瘤比以前假设的。
DNA amplifications are important mechanisms for protooncogene activation. Comparative genomic hybridization (CGH) to metaphase chromosome preparations has revealed amplifications in 10-20% of B-cell lymphomas (B-NHL). We analysed a series of 16 aggressive non-Hodgkin lymphomas by the new approach termed Matrix-CGH (M-CGH) using genomic DNA microarrays as hybridization target. For M-CGH, a dedicated B-cell lymphoma chip was constructed containing 496 genomic targets covering oncogenes, tumor suppressor genes as well as chromosome regions frequently altered in B-NHL. In 10 of 16 samples a total of 15 DNA amplifications were identified. The amplicons included BCL2, REL, CCND1, CCND2, JAK2, FGF4 and MDM2. Four of the 15 amplifications remained undetected by chromosomal CGH. The respective amplicons mapped to bands 2p13, 9p13-p21 and 12q24 and, were confirmed by fluorescence in situ hybridization. Furthermore, for four genomically amplified genes real-time quantitative reverse transcription polymerase chain reaction revealed elevated mRNA expression levels. These data show the superior diagnostic sensitivity of the newly developed diagnostic tool. As only a small portion of the genome (approximately 1.5%) has been analysed by the present DNA array, it is likely that gene amplifications are much more common in aggressive lymphomas than previously assumed.