Kinetic analysis, size profiling, and bioenergetic association of DNA released by selected cell lines in vitro
Kinetic analysis, size profiling, and bioenergetic association of DNA released by selected cell lines in vitro
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DOI：
10.1007/s00018-017-2495-z
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发表时间：
2017-07-01
影响因子：
8
通讯作者：
Pretorius, Piet J.
中科院分区：
文献类型：
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作者：
Aucamp, Janine;Bronkhorst, Abel J.;Pretorius, Piet J.
尽管循环DNA （cirDNA）分析作为一种广泛的病理筛选工具显示出巨大的希望，但许多障碍阻碍了研究到临床实践的快速转化。这与体内环境的内在复杂性直接相关，其中相互关联的细胞反应和假定的DNA来源的复杂系统的影响，创造了任何个体血液中cirDNA的定量和定性特性的看似任意的表示。因此，为了评估体外细胞培养的潜力，以避免在体内研究中遇到的困难，本工作的目的是阐明8种不同细胞系释放的DNA[无细胞DNA (cfDNA)]的特征。这揭示了三种不同形式的cfDNA释放模式和核小体片段的存在以及主动释放形式的DNA，这不仅在每个测试细胞系中都一致观察到，而且在血浆样品中也一致观察到。通过筛选和比较生物能量通量参数，研究cfDNA释放与细胞起源、生长速度和癌症状态的相关性。这些结果表明cfDNA水平与糖酵解之间具有统计学意义的相关性，而cfDNA水平与氧化磷酸化之间没有相关性。此外，还观察到生长速度、癌症状态和对有氧糖酵解的依赖性之间的一些相关性。因此，细胞培养物可以成功地作为闭环模型，替代生物流体样品或与生物流体样品结合使用，这将使对特定细胞类型或DNA起源的关注更加清晰。
Although circulating DNA (cirDNA) analysis shows great promise as a screening tool for a wide range of pathologies, numerous stumbling blocks hinder the rapid translation of research to clinical practice. This is related directly to the inherent complexity of the in vivo setting, wherein the influence of complex systems of interconnected cellular responses and putative DNA sources creates a seemingly arbitrary representation of the quantitative and qualitative properties of the cirDNA in the blood of any individual. Therefore, to evaluate the potential of in vitro cell cultures to circumvent the difficulties encountered in in vivo investigations, the purpose of this work was to elucidate the characteristics of the DNA released [cell-free DNA (cfDNA)] by eight different cell lines. This revealed three different forms of cfDNA release patterns and the presence of nucleosomal fragments as well as actively released forms of DNA, which are not only consistently observed in every tested cell line, but also in plasma samples. Correlations between cfDNA release and cellular origin, growth rate, and cancer status were also investigated by screening and comparing bioenergetics flux parameters. These results show statistically significant correlations between cfDNA levels and glycolysis, while no correlations between cfDNA levels and oxidative phosphorylation were observed. Furthermore, several correlations between growth rate, cancer status, and dependency on aerobic glycolysis were observed. Cell cultures can, therefore, successfully serve as closed-circuit models to either replace or be used in conjunction with biofluid samples, which will enable sharper focus on specific cell types or DNA origins.