DNA strand-breaks induced by the topoisomerase I inhibitor camptothecin in unstimulated human white blood cells.
Camptothecin (CPT) and actinomicyn-induced strand-breaks, repair and apoptosis in unstimulated human blood cells were studied using the DNA comet assay, and electrophoresis of low molecular weight DNA extracts. On the one hand, incubation of G0 leukocytes for 1 h with CPT induced DNA strand-breaks that were observed using the single cell gel electrophoresis technique. On the other hand, internucleosomal DNA fragments were not observed, suggesting that apoptosis had not occurred. DNA-strand-breaks caused by CPT were repaired 24 h after treatment; the migration of DNA fragments was assessed by a reduction in the number of comets. These data strongly suggest that the unexpected clastogenic effect of this topoisomerase I inhibitor is not due to the collision of the cleavage complex with the replication fork, since replication does not occur in G0. In our opinion, this effect could be due instead to the topoisomerase I enzyme being able to bind DNA in the absence of replication, probably in a way that is not strictly related to the progression of the cell cycle. Thus, CPT does not provoke apoptosis in quiescent leukocytes.
Daza
P
P
Department of Cell Biology, Faculty of Biology, Avda. Reina Mercedes 6, 41012 Sevilla, Spain. pdaza@us.es
Torreblanca
J
J
García-Herdugo
G
G
Moreno
F J
FJ
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002166:Camptothecin / Q000494:pharmacology*; D002467:Cell Nucleus / Q000187:drug effects* / Q000201:enzymology / Q000648:ultrastructure; D020552:Comet Assay; D004247:DNA / Q000187:drug effects* / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects* / Q000235:genetics; D004260:DNA Repair / Q000187:drug effects / Q000235:genetics; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D003839:Deoxyadenosines / Q000494:pharmacology; D006801:Humans; D007399:Interphase / Q000187:drug effects / Q000235:genetics; D007962:Leukocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D008297:Male; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D012313:RNA / Q000187:drug effects / Q000235:genetics; D059004:Topoisomerase I Inhibitors
D000972:Antineoplastic Agents, Phytogenic; D003839:Deoxyadenosines; D019384:Nucleic Acid Synthesis Inhibitors; D059004:Topoisomerase I Inhibitors; D003609:Dactinomycin; D012313:RNA; D004247:DNA; D004264:DNA Topoisomerases, Type I; C058120:cordycepin; D002166:Camptothecin