From amplification to gene in thyroid cancer: A high-resolution mapped bacterial-artificial-chromosome resource for cancer chromosome aberrations guides gene discovery after comparative genome hybridization
From amplification to gene in thyroid cancer: A high-resolution mapped bacterial-artificial-chromosome resource for cancer chromosome aberrations guides gene discovery after comparative genome hybridization
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DOI：
10.1086/301973
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发表时间：
1998-08-01
影响因子：
9.8
通讯作者：
Korenberg, JR
中科院分区：
文献类型：
--
作者：
Chen, XN;Knauf, JA;Korenberg, JR
与肿瘤相关的染色体重排为肿瘤发生途径的定义提供了丰富的资源。比较基因组杂交（CGH）识别新基因的能力取决于是否存在合适的标记，而大多数基因组都缺乏这些标记。我们现在报告一种通用方法，将 CGH 数据转化为更高分辨率的基因组克隆数据，然后用于定义位于非整倍体区域的基因。我们使用 CGH 研究了 33 种甲状腺肿瘤 DNA 和两种肿瘤细胞系 DNA。结果显示染色体带 2p21 扩增，2p13、19q13.1 和 1p36 扩增强度较低，1p34、1q42、5q31、5q33-34、9q32-34 和 14q32 扩增强度最低。为了定义扩增的 2p21 区域，构建了 373 个 FISH 定位的 2 号染色体细菌人工染色体 (BAC) 的密集阵列，其中 87 个与肿瘤细胞系杂交。四个 BAC 携带在这些细胞中扩增的基因组 DNA。通过带有侧翼 BAC 的多色 FISH 将最大扩增区域缩小至 3-6 Mb，最小扩增子大小由 420 kb 的重叠群定义。对扩增的 BAC 1D9 进行序列分析，揭示了该基因的一个片段，编码蛋白激酶 C epsilon (PKC epsilon)，随后显示该片段在肿瘤细胞中被扩增并重排。总之，CGH 结合 BAC 的密集图谱资源和大规模测序，直接导致 PKC epsilon 被定义为先前未图谱的参与甲状腺肿瘤发生的候选基因。
Chromosome rearrangements associated with neoplasms provide a rich resource for definition of the pathways of tumorigenesis. The power of comparative genome hybridization (CGH) to identify novel genes depends on the existence of suitable markers, which are lacking throughout most of the genome. We now report a general approach that translates CGH data into higher-resolution genomic-clone data that are then used to define the genes located in aneuploid regions. We used CGH to study 33 thyroid-tumor DNAs and two tumor-cell-line DNAs. The results revealed amplifications of chromosome band 2p21, with less-intense amplification on 2p13, 19q13.1, and 1p36 and with least-intense amplification on 1p34, 1q42, 5q31, 5q33-34, 9q32-34, and 14q32. To define the 2p21 region amplified, a dense array of 373 FISH-mapped chromosome 2 bacterial artificial chromosomes (BACs) was constructed, and 87 of these were hybridized to a tumor-cell line. Four BACs carried genomic DNA that was amplified in these cells. The maximum amplified region was narrowed to 3-6 Mb by multicolor FISH with the flanking BACs, and the minimum amplicon size was defined by a contig of 420 kb. Sequence analysis of the amplified BAC 1D9 revealed a fragment of the gene, encoding protein kinase C epsilon (PKC epsilon), that was then shown to be amplified and rearranged in tumor cells. In summary, CGH combined with a dense mapped resource of BACs;and large-scale sequencing has led directly to the definition of PKC epsilon as a previously unmapped candidate gene involved in thyroid tumorigenesis.