Oxidative stress induces DNA damage and inhibits the repair of DNA lesions induced by N-acetoxy-2-acetylaminofluorene in human peripheral mononuclear leukocytes.
Human mononuclear leukocytes were exposed to prooxidants such as H2O2, phorbol-12-myristate-13-acetate, and 4-nitroquinoline-N-oxide, and the effects on induction of DNA damage and repair were evaluated. ADP ribosylation was activated by prooxidant exposure and the response was bimodal with peaks of activation occurring at about 30 min and 4-5 h. Other evidence for prooxidant-induced DNA damage was provided by nucleoid sedimentation assays. Unscheduled DNA synthesis (UDS) was only slightly induced by prooxidant exposure which suggested that either the DNA lesions were repaired by a short patch mechanism involving little UDS, or the repair process was inhibited by prooxidant exposures, or some combination of both. This point was clarified by the fact that the repair of DNA lesions induced by N-acetoxy-2-acetylaminofluorene, an inducer of large patch DNA repair, was inhibited in a dose-dependent manner by exposure to H2O2 and the inhibition was dependent on ADP ribosylation. In contrast, the repair of DNA strand breaks induced by prooxidant exposures as identified above were complete within about 8 h and the repair was independent of ADP ribosylation. Both ADP ribosylation and N-acetoxy-2-acetylaminofluorene-induced UDS were shown to be up- and down-regulated by the redox state of human mononuclear leukocytes indicating a unique mechanism of cellular control over DNA repair.
Pero
R W
RW
PMI-Strang Clinic, Department of Biochemical Epidemiology, New York, New York 10016.
Anderson
M W
MW
Doyle
G A
GA
Anna
C H
CH
Romagna
F
F
Markowitz
M
M
Bryngelsson
C
C
D015073:2-Acetylaminofluorene; D015112:4-Nitroquinoline-1-oxide / Q000494:pharmacology*; D000099:Acetoxyacetylaminofluorene / Q000494:pharmacology*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D004247:DNA / Q000097:blood / Q000187:drug effects; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D066298:In Vitro Techniques; D007700:Kinetics; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism*; D009600:Nitroquinolines / Q000494:pharmacology*; D010084:Oxidation-Reduction; D011065:Poly(ADP-ribose) Polymerases / Q000097:blood; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D000945:Antigens, Differentiation, T-Lymphocyte; D009600:Nitroquinolines; D015112:4-Nitroquinoline-1-oxide; D000099:Acetoxyacetylaminofluorene; D004247:DNA; D015073:2-Acetylaminofluorene; D006861:Hydrogen Peroxide; D011065:Poly(ADP-ribose) Polymerases; D013755:Tetradecanoylphorbol Acetate