Cancer cell line identification by short tandem repeat profiling:: power and limitations
Cancer cell line identification by short tandem repeat profiling:: power and limitations
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DOI：
10.1096/fj.04-3062fje
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发表时间：
2005-01-01
期刊：
影响因子：
4.8
通讯作者：
Kofler, R
中科院分区：
文献类型：
--
作者：
Parson, W;Kirchebner, R;Kofler, R
癌症细胞系在世界范围内用于生物学研究，数据解释取决于各个细胞系对其原始来源的明确归属。短串联重复序列（STR）分析（DNA指纹）是用于此目的的首选方法;然而，在各种实验条件下细胞系的遗传稳定性并没有得到很好的定义。我们在四种广泛使用的白血病细胞系中测试了长期培养、亚克隆和耐药亚克隆的产生对指纹图谱的影响。DNA指纹图谱保持不变，其中两个（U937和K562）在整个12个月的文化，和绝大多数的亚克隆来源于有限稀释后，长期培养揭示了相同的配置文件，表明高度的稳定性和克隆型同质性。相比之下，另外两个细胞系（CCRF-CEM和Jurkat）在长期培养过程中DNA指纹图谱发生了显著变化。有限稀释亚克隆揭示了广泛的克隆型异质性，亚克隆在多达8个STR基因座与亲本培养物不同。相似的异质性，观察到选择培养产生的耐药性，DNA指纹图谱证明在确定可能的耐药机制的亚克隆。因此，常见的组织培养程序可能会显着影响某些细胞系的指纹图谱，从而使其来源的定义变得困难。
Cancer cell lines are used worldwide in biological research, and data interpretation depends on unambiguous attribution of the respective cell line to its original source. Short-tandem-repeat (STR) profiling ( DNA fingerprinting) is the method of choice for this purpose; however, the genetic stability of cell lines under various experimental conditions is not well defined. We tested the effect of long-term culture, subcloning, and generation of drug-resistant subclones on fingerprinting profiles in four widely used leukemia cell lines. The DNA fingerprinting profile remained unaltered in two of them ( U937 and K562) throughout 12 months in culture, and the vast majority of subclones derived therefrom by limiting dilution after long-term culture revealed the same profile, indicating a high degree of stability and clonotypic homogeneity. In contrast, two other cell lines (CCRF-CEM and Jurkat) showed marked alterations in DNA fingerprinting profiles during long-term culture. Limiting dilution subcloning revealed extensive clonotypic heterogeneity with subclones differing in up to eight STR loci from the parental culture. Similar heterogeneity was observed in subclones generated by selection culture for drug resistance where DNA fingerprinting proved useful in identifying possible resistance mechanisms. Thus, common tissue culture procedures may dramatically affect the fingerprinting profile of certain cell lines and thus render definition of their origin difficult.