Cytotoxicity and glutathione depletion by 1-methyl-2-nitrosoimidazole in human colon cancer cells.
The biological effects of 1-methyl-2-nitrosoimidazole (INO), the 2 electron reduction product of biologically active 1-methyl-2-nitroimidazole, were examined in HT-29 human colon cancer cells by clonogenic assay and glutathione (GSH) determination. INO was very toxic towards HT-29 cells and was equally toxic under aerobic and hypoxic conditions. Cytotoxicity was highly dependent on cell concentration, decreasing as cell concentration increased. INO also resulted in an initial dose-dependent depletion of intracellular GSH which plateaued when the GSH content of INO-treated cells reached approximately 8% of control levels. As was the case for cytotoxicity, the magnitude of GSH depletion by any given INO dose was highly dependent on cell concentration, being greatest at low cell densities. Both toxicity and GSH depletion were more pronounced when cells were exposed in culture medium without the reducing agent, ascorbate. HT-29 cells preincubated with the GSH synthesis inhibitor, buthionine sulfoximine (BSO), to reduce GSH levels to approximately 10% of control levels were more sensitive to subsequent INO exposure. These data suggest that the nitroso- reduction product of 2-nitroimidazoles may be responsible for cytotoxicity and glutathione depletion associated with hypoxic exposure to 2-nitroimidazoles.
Mulcahy
R T
RT
Department of Human Oncology, University of Wisconsin Clinical Cancer Center, Madison 53792.
Gipp
J J
JJ
Ublacker
G A
GA
Panicucci
R
R
McClelland
R A
RA
D019328:Buthionine Sulfoximine; D002470:Cell Survival / Q000187:drug effects; D003110:Colonic Neoplasms / Q000032:analysis / Q000473:pathology; D005978:Glutathione / Q000032:analysis*; D006801:Humans; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D009593:Nitroimidazoles / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000187:drug effects
D009593:Nitroimidazoles; C042596:1-methyl-2-nitroimidazole; D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; D005978:Glutathione