title: The effect of aging on the DNA damage and repair capacity in 2BS cells undergoing oxidative stress.
abstract: Aging is associated with a reduction in the DNA repair capacity under oxidative stress. However, whether the DNA damage and repair capacity can be a biomarker of aging remains controversial. In this study, we demonstrated two cause-and-effect relationships, the one is between the DNA damage and repair capacity and the cellular age, another is between DNA damage and repair capacity and the level of oxidative stress in human embryonic lung fibroblasts (2BS) exposed to different doses of hydrogen peroxide (H2O2). To clarify the mechanisms of the age-related reduction in DNA damage and repair capacity, we preliminarily evaluated the expressions of six kinds of pivotal enzymes involved in the two classical DNA repair pathways. The DNA repair capacity was observed in human fibroblasts cells using the comet assay; the age-related DNA repair enzymes were selected by RT-PCR and then verified by Western blot in vitro. Results showed that the DNA repair capacity was negatively and linearly correlated with (i) cumulative population doubling (PD) levels only in the group of low concentration of hydrogen peroxide treatment, (ii) with the level of oxidative stress only in the group of young PD cells. The mRNA expression of DNA polymerase δ1 decreased substantially in senescent cells and showed negative linear-correlation with PD levels; the protein expression level was well consistent with the mRNA level. Taken together, DNA damage and repair capacity can be a biomarker of aging. Reduced expression of DNA polymerase δ1 may be responsible for the decrease of DNA repair capacity in senescent cells.
authors.affiliation: Department of Clinical Laboratory, Xuanwu Hospital, Capital Medical University, Beijing, 100053, People's Republic of China. wangjinling0224@gmail.com
authors.forename: Jin-Ling
authors.initials: JL
authors.lastname: Wang
authors.forename: Pei-Chang
authors.initials: PC
authors.lastname: Wang
mesh_terms: D015153:Blotting, Western; D016922:Cellular Senescence / Q000502:physiology*; D020552:Comet Assay; D004249:DNA Damage; D004260:DNA Repair / Q000502:physiology*; D005347:Fibroblasts; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D006861:Hydrogen Peroxide