DNA interaction with platinum-based cytostatics revealed by DNA sequencing
|Článek v odborném periodiku
|Časopis / Zdroj
|Fakulta / Pracoviště MU
|DNA; Platinum based cytostatics; Sequencing; Mass spectrometry
|The main mechanism of action of platinum-based cytostatic drugs cisplatin, oxaliplatin and carboplatin - is the formation of DNA cross-links, which restricts the transcription due to the disability of DNA to enter the active site of the polymerase. The polymerase chain reaction (PCR) was employed as a simplified model of the amplification process in the cell nucleus. PCR with fluorescently labelled dideoxynucleotides commonly employed for DNA sequencing was used to monitor the effect of platinum-based cytostatics on DNA in terms of decrease in labeling efficiency dependent on a presence of the DNA-drug cross-link. It was found that significantly different amounts of the drugs - cisplatin (0.21 mu g/mL), oxaliplatin (5.23 mu g/mL), and carboplatin (71.11 mu g/mL) - were required to cause the same quenching effect (50%) on the fluorescent labelling of 50 mu g/mL of DNA. Moreover, it was found that even though the amounts of the drugs was applied to the reaction mixture differing by several orders of magnitude, the amount of incorporated platinum, quantified by inductively coupled plasma mass spectrometry, was in all cases at the level of tenths of mu g per 5 mu g of DNA.