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首页> 美国卫生研究院文献>Oncology Letters >The importance of molecular cytogenetic analysis prior to using cell lines in research: The case of the KG-1a leukemia cell line
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The importance of molecular cytogenetic analysis prior to using cell lines in research: The case of the KG-1a leukemia cell line

机译：在研究中使用细胞系之前进行分子细胞遗传学分析的重要性：KG-1a白血病细胞系的情况

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摘要

KG-1 and its less differentiated subline KG-1a are leukemia cell lines used in research in a number of laboratories. The karyotypes of the two lines were initially identical. In the following years, further analysis revealed that the cell lines had acquired additional karyotypical abnormalities and differed in the presence of certain typical chromosomal rearrangements. To obtain cytogenetic authentication prior to the use of the two cell lines, we analyzed their karyotype by combining DAPI- and CMA-chromosome bandings and a fluorescence in situ hybridization (FISH)-based approach by using BAC clones useful for the identification of chromosome regions of interest. Sequences of the MYC, PLZF, RARA and BCR genes, that are known to play a critical role in leukemogenesis, and certain BAC clones mapped to five known common fragile sites (CFS) were used for the FISH analysis. A telomeric probe (TTAGGG)n and a set of BAC clones were used to characterize the marker chromosome der() that was observed in the cell line KG-1a. The existence of notable differences between the karyotype of the KG-1a cell line previously described, and that described in this study, demonstrate that the use of established cancer cell lines should be preceded by cytogenetic and/or molecular characterization.
机译：KG-1及其分化程度较低的亚系KG-1a是用于许多实验室研究的白血病细胞系。两条系的核型最初是相同的。在接下来的几年中，进一步的分析表明，这些细胞系已经获得了额外的核型异常，并且在某些典型的染色体重排的存在上有所不同。为了在使用这两种细胞系之前获得细胞遗传学认证，我们通过结合使用DAPI和CMA染色体条带以及基于荧光原位杂交（FISH）的方法，通过使用可用于鉴定染色体区域的BAC克隆，分析了它们的核型出于兴趣。已知在白血病发生中起关键作用的MYC，PLZF，RARA和BCR基因的序列，映射到五个已知的常见脆弱位点（CFS）的某些BAC克隆用于FISH分析。使用端粒探针（TTAGGG）n和一组BAC克隆来表征在细胞系KG-1a中观察到的标记染色体der（）。先前描述的KG-1a细胞核型与本研究中描述的核型之间存在显着差异，表明使用已建立的癌细胞系应先进行细胞遗传学和/或分子表征。

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