Oxidative DNA damage and senescence of human diploid fibroblast cells

Qin Chen, Ann Fischer, Joshua D. Reagan, Liang-Jun Yan, Bruce N. Ames

Research output: Contribution to journalArticle

517 Scopus citations

Abstract

Human diploid fibroblast cells cease growth in culture after a finite number of population doublings. To address the cause of growth cessation in senescent IMR-90 human fibroblast cells, we determined the level of oxidative DNA damage by using 8-oxoguanine excised from DNA and 8-oxo-2'-deoxyguanosine in DNA as markers. Senescent cells excise from DNA four times more 8- oxoguanine per day than do early-passage young cells. The steady-state level of 8-oxo-2'-deoxyguanosine in DNA is ≃35% higher in senescent cells than in young cells. Measurement of protein carbonyls shows that senescent cells did not appear to have elevated protein oxidation. To reduce the level of oxidative damage, we cultured cells under a more physiological O2 concentration (3%) and compared the replicative life span to the cells cultured at the O2 concentration of air (20%). We found that cells grown under 3% O2 achieved 50% more population doublings during their lifetime. Such an extension of life span resulted from the delayed onset of senescence and elevation of growth rate and saturation density of cells at all passages. The spin-trapping agent α-phenyl-t-butyl nitrone (PBN), which can act as an antioxidant, also effectively delayed senescence and rejuvenated near senescent cells. The effect is dose-dependent and is most pronounced for cells at the stage just before entry into senescence. Our data support the hypothesis that oxidative DNA damage contributes to replicative cessation in human diploid fibroblast cells.

Original languageEnglish
Pages (from-to)4337-4341
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number10
DOIs
StatePublished - 9 May 1995

Keywords

  • 8-oxoguanine
  • oxygen tension
  • protein oxidation
  • replicative life span
  • α-phenyl-t-butyl nitrone

Fingerprint Dive into the research topics of 'Oxidative DNA damage and senescence of human diploid fibroblast cells'. Together they form a unique fingerprint.

  • Cite this