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. 2016 Jan;1863(1):157-65.
doi: 10.1016/j.bbamcr.2015.10.022. Epub 2015 Oct 31.

Characterization of the cell-free DNA released by cultured cancer cells

Abel Jacobus Bronkhorst  1 , Johannes F Wentzel  2 , Janine Aucamp  3 , Etresia van Dyk  3 , Lissinda du Plessis  2 , Piet J Pretorius  3
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Free article

Characterization of the cell-free DNA released by cultured cancer cells

Abel Jacobus Bronkhorst et al. Biochim Biophys Acta. 2016 Jan.
Free article

Abstract

The most prominent factor that delays the translation of cell-free DNA (cfDNA) analyses to clinical practice is the lack of knowledge regarding its origin and composition. The elucidation of the former is complicated by the seemingly random fluctuation of quantitative and qualitative characteristics of cfDNA in the blood of healthy and diseased individuals. Besides methodological discrepancies, this could be ascribed to a web of cellular responses to various environmental cues and stressors. Since all cells release cfDNA, it follows that the cfDNA in the blood of cancer patients is not only representative of tumor derived DNA, but also of DNA released by healthy cells under different conditions. Additionally, cfDNA released by malignant cells is not necessarily just aberrant, but likely includes non-mutated chromosomal DNA fragments. This may cause false positive/negative results. Although many have acknowledged that this is a major problem, few have addressed it. We propose that many of the current stumbling blocks encountered in in vivo cfDNA studies can be partially circumvented by in vitro models. Accordingly, the purpose of this work was to evaluate the release of cfDNA from cultured cells and to gauge its potential use for elucidating the nature of cfDNA. Results suggest that the occurrence of cfDNA is not a consequence of apoptosis or necrosis, but primarily a result of actively secreted DNA, perhaps in association with a protein complex. This study demonstrates the potential of in vitro cell culture models to obtain useful information about the phenomenon of cfDNA.

Keywords: Apoptosis; Cell-free DNA (cfDNA); Flow cytometry; Necrosis; Osteosarcoma.

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