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采用光谱核型分析对膀胱癌细胞系BK-10进行分子细胞遗传学分析。

Molecular cytogenetic analysis of the bladder carcinoma cell line BK-10 by spectral karyotyping.

作者信息

Padilla-Nash H M, Nash W G, Padilla G M, Roberson K M, Robertson C N, Macville M, Schröck E, Ried T

机构信息

Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.

出版信息

Genes Chromosomes Cancer. 1999 May;25(1):53-9. doi: 10.1002/(sici)1098-2264(199905)25:1<53::aid-gcc8>3.0.co;2-t.

Abstract

The bladder cancer cell line BK-10 was established from a grade III-IV transitional cell carcinoma (TCC). BK-10 is near-tetraploid (+/-4n) and consists of two subclones with 20-25 structural aberrations. Here we report the cytogenetic analysis of BK-10 by G-banding, spectral karyotyping (SKY), and FISH. SKY refers to the hybridization of 24 differentially labeled chromosome painting probes and the simultaneous visualization of all human chromosomes using spectral imaging. SKY enabled us to confirm 12 markers in BK-10 previously described by G-banding, redefine 11 aberrations, and detect 4 hidden chromosomal rearrangements, 2 of which had been identified as normal or deleted copies of chromosome 20 and 1 as a normal chromosome 3. Twenty out of 21 translocations identified were unbalanced. FISH analysis of BK-10 using chromosome arm-specific paints, centromere probes, and oncogene/tumor suppressor gene-specific probes revealed a deletion of CDKN2A (p16) in all copies of chromosome 9, a low-level amplification of MYC (five copies), and loss of one copy of TP53; detected the presence of the Y chromosome in a hidden translocation; and detected four copies of ERBB-2. A probe set for BCR and ABL verified breakpoints for all translocations involving chromosomes 9 and 22. A new karyotype presentation, "SKY-gram," is introduced by combining data from G-banding, SKY, and FISH analysis. This study demonstrates the approach of combining molecular cytogenetic techniques to characterize fully the multiple complex chromosomal rearrangements found in the bladder cancer cell line BK-10, and to refine the chromosomal breakpoints for all translocations.

摘要

膀胱癌细胞系BK - 10源自III - IV级移行细胞癌(TCC)。BK - 10接近四倍体(±4n),由两个具有20 - 25个结构畸变的亚克隆组成。在此,我们报告通过G显带、光谱核型分析(SKY)和荧光原位杂交(FISH)对BK - 10进行的细胞遗传学分析。SKY是指24种差异标记的染色体涂染探针的杂交,并使用光谱成像同时可视化所有人类染色体。SKY使我们能够确认BK - 10中先前通过G显带描述的12个标记,重新定义11个畸变,并检测到4个隐藏的染色体重排,其中2个被鉴定为20号染色体的正常或缺失拷贝,1个为正常的3号染色体。在鉴定出的21种易位中,有20种是不平衡的。使用染色体臂特异性涂染剂、着丝粒探针以及癌基因/肿瘤抑制基因特异性探针对BK - 10进行FISH分析,结果显示9号染色体的所有拷贝中CDKN2A(p16)缺失、MYC低水平扩增(5个拷贝)以及TP53缺失一个拷贝;检测到在一个隐藏的易位中有Y染色体存在;并检测到4个ERBB - 2拷贝。用于BCR和ABL的探针组验证了所有涉及9号和22号染色体的易位的断点。通过整合G显带、SKY和FISH分析的数据,引入了一种新的核型呈现方式“SKY图”。本研究展示了结合分子细胞遗传学技术的方法,以全面表征在膀胱癌细胞系BK - 10中发现的多种复杂染色体重排,并细化所有易位的染色体断点。

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