The effect of butylated hydroxytoluene on the chromosomal damage induced by bleomycin in Chinese hamster ovary cells.
The effect of butylated hydroxytoluene (BHT) on chromosomal damage induced by bleomycin (BLM) in CHO cells was studied. Treatments were performed in cells at quiescent state (90/95% at G0-G1), 2 h after subculture (G1) or 3 h before fixation (G2). Cells were treated for 20 min with BLM plus BHT and subsequently incubated in the presence of BHT until fixation. Results were compared with those obtained from untreated and DMSO-treated controls, from treatments with BLM or BHT alone, from treatments with BLM followed by treatment with DMSO until fixation, and from treatments with BLM plus BHT for 20 min without post-treatment with BHT. BLM induced chromatid- and chromosome-type aberrations in cells treated at G0/G1 or G1 and chromatid-type aberrations in cells treated at the G2 stage. Post-treatment with BHT strongly decreased the frequency of chromosome- but not of chromatid-type aberrations in G0/G1 and G1 and of chromatid-type aberrations in G2. These results are explained assuming that chromosome-type aberrations induced at G0/G1 and G1, and chromatid-type aberrations induced at G2 are originated by the induction of double-strand breaks by BLM through the formation of free radicals. Thus, the observed effect of BHT post-treatment could be considered as evidence that chromosome aberrations are induced by BLM following a two-step mechanism. On the other hand, it is necessary to differentiate between chromatid-type aberrations induced by BLM at G0/G1 and those produced by G2 treatment on the basis of replication errors for the former and DNA repair errors for the latter. In addition, the induction of chromatid-type aberrations by BHT itself at G0/G1 must be taken into account. As BHT acts as an S-dependent agent, chromatid-type aberrations observed after treatment with BLM and BHT in G0/G1 could arise from single-strand breaks induced by BLM and DNA primary lesions induced by BHT.
Grillo
C A
CA
Centro de Investigaciones en Genética Básica y Aplicada (CIGEBA), Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Argentina.
Dulout
F N
FN
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D001761:Bleomycin / Q000494:pharmacology*; D002084:Butylated Hydroxytoluene / Q000494:pharmacology*; D016466:CHO Cells / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002869:Chromosome Aberrations; D006224:Cricetinae; D004249:DNA Damage / Q000187:drug effects*; D004357:Drug Synergism; D016193:G1 Phase / Q000187:drug effects / Q000235:genetics; D016195:G2 Phase / Q000187:drug effects / Q000235:genetics; D016192:Resting Phase, Cell Cycle / Q000187:drug effects / Q000235:genetics
D000964:Antimetabolites, Antineoplastic; D000975:Antioxidants; D001761:Bleomycin; D002084:Butylated Hydroxytoluene