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Cisplatin generates superoxide anion by interaction with DNA in a cell-free system
H Masuda1, T Tanaka, U Takahama
1Department of Surgery, Kyushu Dental College, Japan.
Biochemical and Biophysical Research Communications
|September 15, 1994
Summary
Cisplatin (DDP) interacts with DNA to produce superoxide anion (O2-) and hydroxyl radicals (OH.). This was confirmed by chemiluminescence assays and radical scavengers, indicating DDP
Area of Science:
- Biochemistry
- Chemical Biology
- Pharmacology
Background:
- Cisplatin (cis-Diamminedichloroplatinum (II), DDP) is a widely used chemotherapy drug.
- The precise mechanisms of DDP-induced cytotoxicity, particularly its interaction with DNA and reactive oxygen species (ROS) generation, require further elucidation.
Purpose of the Study:
- To investigate the generation of reactive oxygen species (ROS), specifically superoxide anion (O2-), by Cisplatin (DDP) upon interaction with calf thymus DNA (ct-DNA).
- To explore the potential involvement of hydroxyl radicals (OH.) in this DDP-DNA interaction process.
Main Methods:
- Chemiluminescence assay using a Cypridina luciferin analog (CLA) to detect O2- generation.
- Investigating the effect of varying concentrations of DDP, ct-DNA, and CLA on chemiluminescence intensity.
- Utilizing radical scavengers, including superoxide dismutase (SOD), ascorbic acid, mannitol, ethanol, and formic acid, to identify the types of ROS produced.
Main Results:
- Cisplatin (DDP) demonstrated the ability to generate superoxide anion (O2-) when incubated with calf thymus DNA (ct-DNA) in a phosphate-buffered saline (PBS) solution.
- The intensity of the chemiluminescence signal (CCLA) increased proportionally with higher concentrations of DDP, ct-DNA, or CLA.
- Superoxide dismutase (SOD) and ascorbic acid completely inhibited the DDP-induced chemiluminescence, confirming O2- generation. Hydroxyl radical scavengers partially suppressed the signal, suggesting OH. production as well.
Conclusions:
- Cisplatin (DDP) directly induces the production of superoxide anion (O2-) through its interaction with DNA in a cell-free system.
- The findings suggest that Cisplatin (DDP) may also generate hydroxyl radicals (OH.), contributing to its overall reactive oxygen species (ROS) profile.
- This study provides insights into the chemical mechanisms underlying Cisplatin's (DDP) biological activity, highlighting ROS generation as a key factor.