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      Kinetic analysis, size profiling, and bioenergetic association of DNA released by selected cell lines in vitro.

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          Abstract

          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.

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          Author and article information

          Journal
          Cell. Mol. Life Sci.
          Cellular and molecular life sciences : CMLS
          Springer Nature America, Inc
          1420-9071
          1420-682X
          July 2017
          : 74
          : 14
          Affiliations
          [1 ] Human Metabolomics, North-West University, Hoffman Street, Potchefstroom, 2520, South Africa. aucampj@telkomsa.net.
          [2 ] Human Metabolomics, North-West University, Hoffman Street, Potchefstroom, 2520, South Africa.
          Article
          10.1007/s00018-017-2495-z
          10.1007/s00018-017-2495-z
          28315952
          bf82f08e-9e86-406a-a415-3f57d561dfef
          History

          Circulating DNA,Aerobic glycolysis,Bioenergetics flux parameters,Cell cultures,Cell-free DNA

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