GC-Rich DNA Fragments and Oxidized Cell-Free DNA Have Different Effects on NF-kB and NRF2 Signaling in MSC
It has been established that cell-free DNA circulating in the bloodstream affects cells. The characteristics of cfDNA depend on the physiological state of the organism. As we showed previously, diseases can cause either GC-enrichment of the cell-free DNA
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Vasilina A. Sergeeva, Svetlana V. Kostyuk, Elizaveta S. Ershova, Elena M. Malinovskaya, Tatiana D. Smirnova, Larisa V. Kameneva, and Natalia N. Veiko
Abstract
It has been established that cell-free DNA circulating in the bloodstream affects cells. The characteristics of cfDNA depend on the physiological state of the organism. As we showed previously, diseases can cause either GC-enrichment of the cell-free DNA pool or its oxidation. Thus, in cases of cerebral atherosclerosis, heart attack and rheumatic arthritis the cellfree DNA pool is GC-enriched and, in the case of cancer, both GC-enriched and oxidized. Herein we investigated the time-dependent effect of oxidized and GC-rich cell-free DNA on NF-kB and NRF2 signaling pathways in human mesenchymal stem cells and showed that they affect cells in different ways. Oxidized DNA drastically increases expression of NRF2 in a short period of time, but the effect does not last long. GC-rich DNA causes a prolonged increase in mRNA levels of NF-kB and NRF2 which lasts 48 and 24 h, respectively. Keywords
Cell-free DNA • MSCs • Oxidized cell-free DNA • GC-rich cell-free DNA
Introduction
V.A. Sergeeva (*) • S.V. Kostyuk • E.S. Ershova • E.M. Malinovskaya • T.D. Smirnova • L.V. Kameneva N.N. Veiko “Research Centre for Medical Genetics”, Federal State Budgetary Institution, Moskvorechie str., 1, Moscow 115478, Russia e-mail: [email protected]
It is a well-known fact that cell-free DNA (cfDNA) circulates throughout the bloodstream, affecting the cells. We investigated various potential biological functions of cfDNA and demonstrated that a GC-rich cfDNA or oxidized cfDNA is a stress signal. As shown previously, a pool of cfDNA consists of GC-rich ribosomal DNA in the serum of patients with some diseases (cere-
© Springer International Publishing Switzerland 2016 P.B. Gahan et al. (eds.), Circulating Nucleic Acids in Serum and Plasma – CNAPS IX, Advances in Experimental Medicine and Biology 924, DOI 10.1007/978-3-319-42044-8_21
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bral atherosclerosis, ischaemic heart disease, heart attack, rheumatic arthritis, cancer) and in some cases, the cfDNA is oxidized (cancer) (Loseva et al. 2012; Ermakov et al. 2013). Our goal was to investigate the effect of these DNA fragments on major signaling pathways.
Statistical Processing of the results was performed using a standard Statgraphics software package. Mean values were compared using the t – test. All reported results were reproduced at least three times as independent biological replicates.
Materials and Methods
Results and Discussion
Cells Human mesenchymal stem cells (MSCs) derived from breast adipose tissue were cultivated at 37.°C in AmnioMax C-100 Basal Medium (“Gibco”) containing AmnioMax Supplement C-100, 20 mmol/L HEPES (“Pan/ Eco”) and antibiotics. MSCs were characterized by a number of standard markers (the cultured MSCs expressed major histocompatibility complex molecules (HLA-ABC+) and adhesion molecules (CD44+, CD54 (low), CD90+, CD106+, CD29+, CD49b (low), and CD105); how
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