Abstract
Purpose
In the following, the cellular and molecular mechanism of cytotoxicity induced by prodrug dacarbazine toward the isolated rat hepatocytes was studied.
Method
Accelerated cytotoxicity screening technique (ACMS) was used to perform this study.
Result
Addition of dacarbazine to isolated rat hepatocytes resulted in reactive oxygen species (ROS) formation, and lysosomal membrane leakiness before hepatocyte lysis occurred. Hepatocyte ROS generation was inhibited by desferoxamine (a ferric chelator). Cytotoxicity was prevented by antioxidants or ROS scavengers (mannitol or dimethylsulfoxide), cytochorome P450 inhibitors (phenylimidazole, diphenyliodonium chloride, 4-methylpyrazole, and benzylimidazole). In addition to lysosomal damage, dacarbazine caused hepatocyte protease activation and cell proteolysis.
Conclusion
Dacarbazine cytotoxicity is associated with ROS (H2O2, O •−2 ) generation. It is suggested that H2O2 could cross the lysosomal membrane, react with lysosomal Fe2+ to form hydroxyl radical (Haber-Weiss reaction) which is the major cause of lysosomal membrane leakiness, proteases, and other digestive enzymes' release and finally the cell death.
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Abbreviations
- DTIC:
-
5-(3, 3-Dimethyl-1-triazeno)-imidazole-4-carboxamide
- MTIC:
-
5-(3-Monomethyl-1-triazeno)-imidazole-4-carboxamide
- AIC:
-
Aminoimidazole carboxamide
- ROS:
-
Reactive oxygen species
- SD:
-
Standard deviation
- ANOVA:
-
Analysis of variance
- DMSO:
-
Dimethylsulfoxide
- DCF:
-
Dichlorofluorescein
- HEPES:
-
(2-Hydroxyethyl)-1-piperazine-ethansulfonic acid)
- h:
-
Hour
- BHA:
-
Butylated hydroxyanisole
- BHT:
-
Butylated hydroxy toluene
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Pourahmad, J., Amirmostofian, M., Kobarfard, F. et al. Biological reactive intermediates that mediate dacarbazine cytotoxicity. Cancer Chemother Pharmacol 65, 89–96 (2009). https://doi.org/10.1007/s00280-009-1007-8
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DOI: https://doi.org/10.1007/s00280-009-1007-8