Detection of multiple gene amplifications in glioblastoma multiforme using array-based comparative genomic hybridization
Detection of multiple gene amplifications in glioblastoma multiforme using array-based comparative genomic hybridization
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DOI：
10.1038/labinvest.3780280
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发表时间：
2001-05-01
影响因子：
5
通讯作者：
Ng, HK
中科院分区：
文献类型：
--
作者：
Hui, ABY;Lo, KW;Ng, HK
我们使用了一种新的基因组芯片的方法来调查扩增的癌基因在整个基因组的多形性胶质母细胞瘤（GBM）。基于阵列的比较基因组杂交（阵列CGH）允许同时检查在各种人类癌症中通常扩增的58个癌基因/扩增子。人类癌症中多种癌基因的扩增可以在一个实验中快速确定。肿瘤DNA和正常对照DNA通过缺口平移分别用绿色和红色标记的核苷酸标记。代替常规比较基因组杂交（CGH）中与正常中期染色体杂交，将混合荧光标记DNA的探针应用于由58个靶癌基因的P1、PAC和BAC克隆组成的基因组阵列模板。通过使用从正常个体制备的测试和参考DNA进行一系列独立阵列CGH来建立测量偏差的基线。在本研究中，我们用CGH和阵列CGH检查了14个GBM（7个细胞系和7个肿瘤），以揭示与这种癌症相关的特定癌基因。在癌基因/扩增子CDK 4、GLI、MYCN、MYC、MDM 2和PDGFRA上鉴定出高水平扩增。在PIK 3CA（64.3%）、EGFR（57.1%）、CSE 1 L（57.1%）、NRAS（50%）、MYCN（42.9%）、FGR（35.7%）、ESR（35.7%）、PGY 1（35.7%）和D17 S167（35.7%）上检测到的增益频率最高。提示这些基因可能参与GEM的发生。
We have used a new method of genomic microarray to investigate amplification of oncogenes throughout the genome of glioblastoma multiforme (GBM). Array-based comparative genomic hybridization (array CGH) allows for simultaneous examination of 58 oncogenes/amplicons that are commonly amplified in various human cancers. Amplification of multiple oncogenes in human cancers can be rapidly determined in a single experiment. Tumor DNA and normal control DNA were labeled by nick translation with green- and red-tagged nucleotides, respectively. Instead of hybridizing to normal metaphase chromosomes in conventional comparative genomic hybridization (CGH), the probes of the mixed fluorescent labeled DNA were applied to genomic array templates comprised of P1, PAC, and BAC clones of 58 target oncogenes. The baseline for measuring deviations was established by performing a series of independent array CGH using test and reference DNA made from normal individuals. In the present study, we examined fourteen GBMs (seven cell lines and seven tumours) with CGH and array CGH to reveal the particular oncogenes associated with this cancer. High-level amplifications were identified on the oncogenes/amplicons CDK4, GLI, MYCN, MYC, MDM2, and PDGFRA. The highest frequencies of gains were detected on PIK3CA (64.3%), EGFR (57.1%), CSE1L (57.1%), NRAS (50%), MYCN (42.9%), FGR (35.7%), ESR (35.7%), PGY1 (35.7%), and D17S167 (35.7%). These genes are suggested to be involved in the GEM tumorigenesis.