Abstract
Size and termini of cell-free DNA molecules circulating in blood plasma and being bound with blood cell surface of healthy females and untreated breast cancer patients were investigated. The size and concentration of circulating blood DNA were analyzed by Agilent 2100 Bioanalyser TM and TaqMan PCR. The termini of circulating DNA were examined by ligation using biotinylated double-stranded oligonucleotide adapters with random 1–3 b overhangs of both chains and subsequent quantification by PCR. Short (180 bp) and longer (>8 kbp) DNA fragments were found in cell free DNA from both groups, but short were less represented in primary breast cancer patient plasma. Predominantly high molecular weight DNA was found in cell surface bound DNA both in healthy females and breast cancer patients with a minor fraction of short fragments. Heterogeneous DNA molecules with diverse 5′- and 3′- protruding as well as blunt ends were found both in plasma DNA and cell bound DNA from healthy individuals. Cell surface bound DNA from breast cancer patients mainly contains 5′-protruding ends, whereas 5′- and 3′-protruding ends are equally presented in cell free DNA from these patients. The data obtained obviously reflect over-representation of specific nucleases in breast cancer.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Bryzgunova OE, Laktionov PP (2014) Generation of blood circulating DNAs: sources, features of struction and circulation. Biochem Mosc Suppl Ser B Biomed Chem 8:203–219. doi:10.1134/S1990750814030020
Bryzgunova O, Bondar A, Morozkin E et al (2011) A reliable method to concentrate circulating DNA. Anal Biochem 408:354–356
Deligezer U, Yaman F, Erten N et al (2003) Frequent copresence of methylated DNA and fragmented nucleosomal DNA in plasma of lymphoma patients. Clin Chim Acta 335:89–94
Didenko VV, Ngo H, Baskin DS (2003) Early necrotic DNA degradation: presence of blunt-ended DNA breaks, 3′ and 5′ overhangs in apoptosis, but only 5′ overhangs in early necrosis. Am J Pathol 162:1571–1578
Suzuki N, Kamataki A, Yamaki J et al (2008) Characterization of circulating DNA in healthy human plasma. Clin Chim Acta 387:55–58
Tamkovich SN, Bryzgunova OE, Rykova EY et al (2005) Circulating nucleic acids in blood of healthy male and female donors. Clin Chem 51:1317–1319
Wu TL, Zhang D, Chia JH et al (2002) Cell-free DNA: measurement in various carcinomas and establishment of normal reference range. Clin Chim Acta 321:77–87
Acknowledgements
The present work was supported by the grant from the Russian Science Support Foundation (13-04-01460).
Conflicts of Interest
The authors declare no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer International Publishing Switzerland
About this paper
Cite this paper
Tamkovich, S.N., Kirushina, N.A., Voytsitskiy, V.E., Tkachuk, V.A., Laktionov, P.P. (2016). Features of Circulating DNA Fragmentation in Blood of Healthy Females and Breast Cancer Patients. In: Gahan, P., Fleischhacker, M., Schmidt, B. (eds) Circulating Nucleic Acids in Serum and Plasma – CNAPS IX. Advances in Experimental Medicine and Biology, vol 924. Springer, Cham. https://doi.org/10.1007/978-3-319-42044-8_10
Download citation
DOI: https://doi.org/10.1007/978-3-319-42044-8_10
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-42042-4
Online ISBN: 978-3-319-42044-8
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)