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Biological reactive intermediates that mediate dacarbazine cytotoxicity

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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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Authors and Affiliations

  1. Faculty of Pharmacy, Shaheed Beheshti University of Medical Sciences, Tehran, P.O. Box 14155-6153, Iran

    Jalal Pourahmad, Marzieh Amirmostofian, Farzad Kobarfard & Jafar Shahraki

Authors
  1. Jalal Pourahmad
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  2. Marzieh Amirmostofian
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  3. Farzad Kobarfard
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  4. Jafar Shahraki
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Correspondence to Jalal Pourahmad.

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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

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