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首页> 外文期刊>Cellular Oncology: Analytical Cellular Pathology >Improved Sensitivity in Comparative Genomic Hybridization Analysis of DNA Heteroploid Cell Mixtures after Pre-Enrichment of Subpopulations by Fluorescence Activated Cell Sorting
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Improved Sensitivity in Comparative Genomic Hybridization Analysis of DNA Heteroploid Cell Mixtures after Pre-Enrichment of Subpopulations by Fluorescence Activated Cell Sorting

机译：通过荧光激活细胞分选预富集亚群后DNA异倍体细胞混合物的比较基因组杂交分析中的灵敏度提高

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摘要

Cytogenetic analysis of solid tumors with comparative genomic hybridization (CGH) is hampered by the dilution of DNA from individual tumor subpopulations with DNA from other cells. We investigated to what extent this dilution effect can be alleviated using fluorescence activated cell sorting (flow sorting) of experimental DNA heteroploid cell mixtures prior to CGH. From mixtures of normal lymphocytes with triploid K‐562 cells the individual components were sorted according to stemline DNA content and processed by CGH in comparison with pure K‐562 samples and the original mixtures. Compared with 30 autosome copy number imbalances found in pure K‐562 samples, a mixture with 32% K‐562 cells showed 16 imbalancies, and none were detected in mixtures with 13% or 5% K‐562 cells. In contrast, 29, 22 and 23 imbalances were detected in K‐562 nuclei sorted from the 32%, 13% and 5% mixtures, respectively. This indicate that CGH analysis of flow sorted DNA aneuploid subpopulations enables a specific cytogenetic analysis of the individual subclones in a DNA heteroploid cell population.
机译：用比较基因组杂交（CGH）对实体瘤进行细胞遗传学分析时，会用来自其他肿瘤细胞的DNA稀释来自各个肿瘤亚群的DNA造成阻碍。我们研究了在CGH之前使用实验性DNA异倍体细胞混合物的荧光激活细胞分选（流动分选）可以在多大程度上减轻这种稀释效果。从正常淋巴细胞与三倍体K-562细胞的混合物中，根据干系DNA含量对各个成分进行分类，并与纯K-562样品和原始混合物相比，由CGH处理。与纯K-562样品中发现的30种常染色体拷贝数失衡相比，具有32％K-562细胞的混合物显示16种不平衡，而在具有13％或5％K-562细胞的混合物中未检测到任何缺陷。相比之下，在分别由32％，13％和5％的混合物分类的K-562核中检测到29、22和23个失衡。这表明对流分类的DNA非整倍体亚群进行CGH分析可以对DNA异倍体细胞群体中的各个亚克隆进行特定的细胞遗传学分析。

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