The comet assay differentiates efficiently and rapidly between genotoxins and cytotoxins in quiescent cells.
Our main aim was to establish the efficiency of the single cell electrophoresis technique for differentiating between drugs that bind DNA and those that do not. The alkaline comet assay was used to test the responses of human leukocytes (quiescent cells) to damage induced by reportedly genotoxic and reportedly cytotoxic agents. Incubation of G0 leukocytes for 1 h with the genotoxic agents camptothecin and actinomycin C provoked DNA migration, observed as comet figures. On the other hand, when cells were treated with the cytotoxic agents cordycepin, fluorodeoxyuridine and puromycin, the leukocyte nuclei were indistinguishable from those of untreated cells. In addition, we have developed a rapid method using non-proliferating cells that requires neither culture nor lymphocyte isolation. This method promises to be useful as a rapid in vitro screening assay.
Daza
P
P
Department of Cell Biology, Faculty of Biology, Avda. Reina Mercedes 6, 41012 Sevilla, Spain. pdaza@us.es
Torreblanca
J
J
Moreno
F J
FJ
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002166:Camptothecin / Q000494:pharmacology; D020552:Comet Assay; D004247:DNA / Q000187:drug effects* / Q000378:metabolism; D003609:Dactinomycin / Q000031:analogs & derivatives* / Q000494:pharmacology; D003839:Deoxyadenosines / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005467:Floxuridine / Q000494:pharmacology; D006801:Humans; D007962:Leukocytes / Q000187:drug effects* / Q000502:physiology; D008297:Male; D009153:Mutagens / Q000494:pharmacology*; D011691:Puromycin / Q000494:pharmacology
D000964:Antimetabolites, Antineoplastic; D000970:Antineoplastic Agents; D003839:Deoxyadenosines; D004791:Enzyme Inhibitors; D009153:Mutagens; D005467:Floxuridine; C030013:cactinomycin; D003609:Dactinomycin; D011691:Puromycin; D004247:DNA; C058120:cordycepin; D002166:Camptothecin