DNA double-strand break repair is impaired in presenescent Syrian hamster fibroblasts

• DOI: 10.1186/s12867-015-0046-4
• Publication Date: 2015-10-13T02:03:58Z
• Authors: Ljudmila Solovjeva; Denis Firsanov; Anastasia Vasilis...; Vadim Chagin; Nadezhda Pleskach; Andrey Kropotov; Maria Svetlova
• Keywords: Immunofluorescence

Studies of DNA damage response are critical for the comprehensive understanding age-related changes in cells, tissues and organisms. Syrian hamster cells halt proliferation become presenescent after several passages standard conditions cultivation due to what is known as «culture stress». Using proliferating young non-dividing primary cultures fibroblasts, we defined their action radiomimetic drug bleomycin (BL) that induces double-strand breaks (DSBs). The effect was estimated by immunoblotting immunofluorescence microscopy using antibody phosphorylated histone H2AX (gH2AX), which generally accepted a DSB marker. At all stages cell cycle, both demonstrated variability number gH2AX foci per nucleus. focus induction found be independent from BL-hydrolase expression. Some differences repair process between BL-treated were observed: (1) kinetics loss G0 fibroblasts culture faster than prematurely stopped dividing; (2) characterized slower recruitment proteins 53BP1, phospho-DNA-PK phospho-ATM focal sites, while rate ATM/ATR substrate accumulation same cells. Our results demonstrate an impairment aged comparison with suggesting BL therapy.