Inhibition of etoposide-induced DNA damage and cytotoxicity in L1210 cells by dehydrogenase inhibitors and other agents.
The mechanism of action of 4'-demethylepipodophyllotoxin-9-(4,6-O-ethylidene-beta-D-glucopyra noside) (VP-16), an important antitumor agent, is unclear. There is evidence that DNA may be the target of action because VP-16 causes single-strand and double-strand breaks in DNA and produces cytotoxicity over a similar dose range. We have hypothesized that an enzyme system, such as dehydrogenase, catalyzes an oxidation-reduction reaction involving the pendant phenolic group which forms an active metabolite that causes the DNA damage and cytotoxicity. To test our hypothesis, we investigated the effect of disulfiram, an aldehyde dehydrogenase inhibitor, and its metabolite, diethyldithiocarbamate, on VP-16-induced DNA damage in L1210 cells. Using the alkaline elution technique to assay DNA damage, we found that disulfiram and diethyldithiocerbamate inhibited VP-16-induced single-strand breaks. Both compounds were also capable of significantly reducing VP-16-induced cytotoxicity. Oxalic acid, pyrophosphate, and malonic acid, competitive inhibitors of succinate dehydrogenase, and the naturally occurring dehydrogenase substrates, succinic acid, beta-glycerophosphate, and isocitric acid, also blocked the effects of VP-16. Free-radical scavengers were also studied. While sodium benzoate was particularly effective in preventing drug-induced DNA damage and cytotoxicity, a number of other scavengers were not. Our data are consistent with the hypothesis that VP-16 is activated by an enzyme such as a dehydrogenase which transforms it into an active intermediate resulting in DNA damage and, consequently, cell death.
D000444:Aldehyde Dehydrogenase; D000445:Aldehyde Oxidoreductases / Q000037:antagonists & inhibitors; D000818:Animals; D001565:Benzoates / Q000494:pharmacology; D019817:Benzoic Acid; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism*; D004221:Disulfiram / Q000494:pharmacology*; D004050:Ditiocarb / Q000494:pharmacology*; D005047:Etoposide / Q000037:antagonists & inhibitors* / Q000627:therapeutic use; D007939:Leukemia L1210 / Q000188:drug therapy* / Q000235:genetics; D051379:Mice; D009249:NADP / Q000494:pharmacology; D011034:Podophyllotoxin / Q000031:analogs & derivatives*; D013859:Thiocarbamates / Q000494:pharmacology*
D001565:Benzoates; D013859:Thiocarbamates; D009249:NADP; D005047:Etoposide; D019817:Benzoic Acid; D004247:DNA; D004050:Ditiocarb; D000445:Aldehyde Oxidoreductases; D000444:Aldehyde Dehydrogenase; D011034:Podophyllotoxin; D004221:Disulfiram