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

BMC Molecular Biology

Research Article

Anastasia Vasilishina¹  Andrey Kropotov¹  Vadim Chagin¹  Nadezhda Pleskach¹  Maria Svetlova¹  Ljudmila Solovjeva¹  Denis Firsanov² 

[1] Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretski ave., 194064, Saint Petersburg, Russia;Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretski ave., 194064, Saint Petersburg, Russia;Saint-Petersburg’s State Pediatric Medical University, Ministry of Health of Russian Federation, 2 Litovskaya st., 194100, Saint Petersburg, Russia;

关键词: Premature aging;    Syrian hamster;    Bleomycin;    Double-strand break repair;   

DOI  :  10.1186/s12867-015-0046-4

 received in 2014-10-21, accepted in 2015-09-28,  发布年份 2015

来源: Springer

【 摘 要 】

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

【 授权许可】

CC BY   
© Solovjeva et al. 2015