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THE ISOLATION AND IDENTIFICATION OF CIS-DIAMMINEDICHLOROPLATINUM (II)-DNA ADDUCTS BY ANION-EXCHANGE HPLC AND INDUCTIVELY COUPLED PLASMA MASS-SPECTROMETRY

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The adducts produced by the reaction of cis-diamminedichloroplatinum(II) with DNA have previously been isolated and characterised. These adducts may be measured at the cellular level by immunochemical detection but the accuracy of this assay is dependent on the number of adducts per nucleotide. We have developed a novel assay for cisplatin-DNA adducts, utilising an established method in which platinated DNA is digested to form a mixture of nucleotides and adducts; these are then separated by anion exchange HPLC. The number of cisplatin-DNA adducts is determined by measurement of the platinum content of the HPLC fractions by inductively coupled plasma mass spectrometry. The assay has been validated by cochromatography of purified drug-DNA adducts whose identity has been confirmed by NMR. We describe an application of the assay, namely the measurement of in vitro removal of cisplatin-DNA adducts from calf thymus DNA by cell free extracts derived from tumour cell lines. Adduct removal is dependent on both the amount of extract protein and the duration of the reaction. Almost 70% of adducts are removed from 5mug of DNA (drug:nucleotide ratio 0.08) by 80mug of extract. Other potential applications of the assay are discussed.
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Document Type: Research Article

Affiliations: 1: UNIV GLASGOW, DEPT MED ONCOL,CRC,ALEXANDER STONE BLDG, GARSCUBE ESTATE, GLASGOW G61 1BD, SCOTLAND. SCOTTISH UNIV RES & REACTOR CTR, E KILBRIDE G75 0QU, LANARK, SCOTLAND. TNO, MED BIOL LAB, RIJSWIJK, NETHERLANDS. QUEEN ELIZABETH HOSP, DEPT MED ONCOL, CRC, BIRMINGHAM B15 2TH, W MIDLANDS, ENGLAND. 2: UNIV GLASGOW, DEPT MED ONCOL,CRC,ALEXANDER STONE BLDG, GARSCUBE ESTATE, GLASGOW G61 1BD, SCOTLAND. SCOTTISH UNIV RES & REACTOR CTR, E KILBRIDE G75 0QU, LANARK, SCOTLAND. TNO, MED BIOL LAB, RIJSWIJK, NETHERLANDS. QUEEN ELIZABETH HOSP, DEPT MED ONCOL, CRC, BIRMINGHAM B15 2TH, W MIDLANDS, ENGLAND. 3: UNIV GLASGOW, DEPT MED ONCOL,CRC,ALEXANDER STONE BLDG, GARSCUBE ESTATE, GLASGOW G61 1BD, SCOTLAND. SCOTTISH UNIV RES & REACTOR CTR, E KILBRIDE G75 0QU, LANARK, SCOTLAND. TNO, MED BIOL LAB, RIJSWIJK, NETHERLANDS. QUEEN ELIZABETH HOSP, DEPT MED ONCOL, CRC, BIRMINGHAM B15 2TH, W MIDLANDS, ENGLAND.

Publication date: January 1, 1993

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  • The International Journal of Oncology provides an international forum for the publication of the latest, cutting-edge research in the broad area of oncology and cancer treatment. The journal accepts original high quality works and reviews on all aspects of oncology research including carcinogenesis, metastasis, epidemiology, chemotherapy and viral oncology. Through fair and efficient peer review, the journal is dedicated to publishing top tier research in the field, offering authors rapid publication as well as high standards of copy-editing and production. The International Journal of Oncology is published on a monthly basis in both print and early online.
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