Differences in DNA damage produced by incorporation of 5-aza-2'-deoxycytidine or 5,6-dihydro-5-azacytidine into DNA of mammalian cells.
The effects of 5-aza-2'-deoxycytidine (aza-dCyd) and 5,6-dihydro-5-azacytidine (H2-aza-Cyd) on the integrity of DNA from several mammalian cell lines were compared using the alkaline elution technique. While both compounds have been shown to inhibit DNA methylation, a direct comparison of their effects on DNA structure has not previously been reported. Exposure of L1210 cells to H2-aza-Cyd (1-100 micrograms/ml) and simultaneous labeling with [14C]thymidine for 24 h resulted in the production of single-strand breaks in DNA, which were significantly repaired when cells were incubated in drug-free medium for an additional 24 h. This differed from our previous findings for aza-dCyd, confirmed here in parallel experiments, which showed that this compound produces alkali-labile lesions that persist for 48 h. The DNA effects of both drugs were significantly reduced when cells were prelabeled with [14C]thymidine, indicating that production of DNA lesions requires incorporation of the anomalous base. Studies utilizing pulse-labeled DNA indicated that aza-dCyd has little effect on the rate of DNA elongation, whereas H2-aza-Cyd produced a complete inhibition for at least 6 h after drug removal. The contrasting pattern of DNA damage induced by these compounds in L1210 was also observed in two human lymphoblastoid cells lines, one of which was derived from a patient with xeroderma pigmentosum. We had previously concluded that alkali-labile sites in DNA from aza-dCyd-treated cells probably arise due to the chemical instability of aza-dCyd. In contrast, incorporated H2-aza-Cyd is chemically stable. The single-strand breaks produced in H2-aza-Cyd treated cells were not of the alkali-labile type, and may represent an accumulation of DNA replication fragments and/or intermediates in an excision repair process. Thus, the DNA lesions produced by the two drugs have markedly different characteristics, and H2-aza-Cyd should not be considered to be merely a stable pharmacological congener of aza-dCyd.
D000818:Animals; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D004261:DNA Replication / Q000187:drug effects; D000077209:Decitabine; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007939:Leukemia L1210; D008214:Lymphocytes; D051379:Mice
C013938:5,6-dihydro-5-azacytidine; D000077209:Decitabine; D004247:DNA; D001374:Azacitidine