Increased DNA-binding activity of cis-1,1-cyclobutanedicarboxylatodiammineplatinum(II) (carboplatin) in the presence of nucleophiles and human breast cancer MCF-7 cell cytoplasmic extracts: activation theory revisited.

Abstract

The molecular mechanism of carboplatin [cis-1,1-cyclobutanedicarboxylatodiammineplatinum(II)] activation is still unresolved. We studied the binding of carboplatin to calf thymus DNA in the presence of thiourea, glutathione, and human breast cancer MCF-7 cell cytoplasmic extracts by measurement of DNA-dependent ethidium bromide fluorescence and atomic absorption spectroscopy. After a 96-hr period of reaction, the decrease in the DNA-dependent fluorescence yield of ethidium bromide due to the formation of platinum (Pt)-DNA adducts increased significantly in the presence of thiourea (6-fold) and glutathione (3- to 4-fold) as compared to the controls in the absence of the nucleophiles. There was also a marked elevation in the levels of platinum incorporated into DNA, measured by atomic absorption spectroscopy (2- to 3-fold and 5- to 7-fold for thiourea and glutathione, respectively). More remarkably, the Pt-DNA adducts formed in the presence of cytoplasmic extracts of MCF-7 human breast cancer cells also showed similar results in a dose-related fashion. Carboplatin, therefore, displayed a characteristic increase in DNA binding/damaging in the presence of the very same S-containing nucleophiles that showed the expected quenching effects in the case of cisplatin [cis-diamminedichloroplatinum (II)]. We propose a nucleophile-facilitated release of the active species of carboplatin prior to binding with DNA.

• Authors: G Natarajan, R Malathi, E Holler
• Journal: Biochemical pharmacology (Biochem Pharmacol) Vol. 58 Issue 10 Pg. 1625-9 (Nov 15 1999) ISSN: 0006-2952 [Print] England
• PMID: 10535754 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antineoplastic Agents
• Cell Extracts
• DNA
• Methionine
• Carboplatin
• Ethidium

Topics

• Antineoplastic Agents (metabolism, pharmacology)
• Breast Neoplasms (pathology)
• Carboplatin (metabolism, pharmacology)
• Cell Extracts (pharmacology)
• DNA (drug effects, metabolism)
• Ethidium (metabolism)
• Fluorescence
• Humans
• Methionine (metabolism)
• Tumor Cells, Cultured