Premium
Circulating cell‐free DNA release in vitro: kinetics, size profiling, and cancer‐related gene methylation
Author(s) -
Panagopoulou Maria,
Karaglani Makrina,
Balgkouranidou Ioanna,
Pantazi Chrisoula,
Kolios George,
Kakolyris Stylianos,
Chatzaki Ekaterini
Publication year - 2019
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.28097
Subject(s) - hela , dna methylation , in vitro , apoptosis , microbiology and biotechnology , methylation , dna , biology , population , gene , demethylating agent , cell culture , cell , chemistry , cancer research , gene expression , biochemistry , genetics , medicine , environmental health
Circulating cell‐free DNA (ccfDNA) is a biological entity of great interest due to its potential as liquid biopsy biomaterial carrying clinically valuable information. To better understand its nature, we studied ccfDNA in vitro in two human cancer cell lines MCF‐7 and HeLa. Normalized indexes of ccfDNA per cell population decreased over time of culture but were significantly elevated after exposure to IC 50 doses of the demethylating/apoptotic agent 5‐azacytidine (5‐AZA‐CR). Fragment‐size profiling was indicative of active release, whereas exposure to 5‐AZA‐CR induced the release of additional shorter fragments, indicative of apoptosis. Finally, the methylation profile of a panel of cancer‐specific genes as assessed by quantitative methylation analysis in ccfDNA was identical to the corresponding genomic DNA and followed accurately changes caused by 5‐AZA‐CR. Overall, our in vitro findings support that ccfDNA can be a reliable biosource of clinically relevant information that can be further studied in these cell culture models.