GC-Rich Extracellular DNA Induces Oxidative Stress, Double-Strand DNA Breaks, and DNA Damage Response in Human Adipose-Derived Mesenchymal Stem Cells.
BACKGROUND
Cell free DNA (cfDNA) circulates throughout the bloodstream of both healthy people and patients with various diseases. CfDNA is substantially enriched in its GC-content as compared with human genomic DNA.


RESULTS
Exposure of haMSCs to GC-DNA induces short-term oxidative stress (determined with H2DCFH-DA) and results in both single- and double-strand DNA breaks (comet assay and γH2AX, foci). As a result in the cells significantly increases the expression of repair genes (BRCA1 (RT-PCR), PCNA (FACS)) and antiapoptotic genes (BCL2 (RT-PCR and FACS), BCL2A1, BCL2L1, BIRC3, and BIRC2 (RT-PCR)). Under the action of GC-DNA the potential of mitochondria was increased. Here we show that GC-rich extracellular DNA stimulates adipocyte differentiation of human adipose-derived mesenchymal stem cells (haMSCs). Exposure to GC-DNA leads to an increase in the level of RNAPPARG2 and LPL (RT-PCR), in the level of fatty acid binding protein FABP4 (FACS analysis) and in the level of fat (Oil Red O).


CONCLUSIONS
GC-rich fragments in the pool of cfDNA can potentially induce oxidative stress and DNA damage response and affect the direction of mesenchymal stem cells differentiation in human adipose-derived mesenchymal stem cells. Such a response may be one of the causes of obesity or osteoporosis.
Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Mosskvorechie Street 1, Moscow 115478, Russia.
Svetlana
S
Kostyuk
Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Mosskvorechie Street 1, Moscow 115478, Russia.
Tatiana
T
Smirnova
Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Mosskvorechie Street 1, Moscow 115478, Russia.
Larisa
L
Kameneva
Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Mosskvorechie Street 1, Moscow 115478, Russia.
Lev
L
Porokhovnik
V.A. Nasonova Research Institute of Rheumatology, Russian Academy of Medical Sciences, Russia.
Anatolij
A
Speranskij
Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Mosskvorechie Street 1, Moscow 115478, Russia.
Elizaveta
E
Ershova
Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Mosskvorechie Street 1, Moscow 115478, Russia.
Sergey
S
Stukalov
Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Mosskvorechie Street 1, Moscow 115478, Russia.
Vera
V
Izevskaya
Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Mosskvorechie Street 1, Moscow 115478, Russia.
Natalia
N
Veiko
D002454:Cell Differentiation; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects*; D004249:DNA Damage / Q000235:genetics*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D018384:Oxidative Stress