An investigation of the sequence-specific interaction of cis-diamminedichloroplatinum(II) and four analogues, including two acridine-tethered complexes, with DNA inside human cells.
The sequence specificity of DNA damage caused by cis-diamminedichloroplatinum(II) (cisplatin) and four analogues in human (HeLa) cells was studied using Taq DNA polymerase and a linear amplification system. The primer extension is inhibited by the drug-DNA adducts, and hence the sites of these lesions can be analyzed on DNA sequencing gels. The repetitive alphoid DNA was used as the target DNA in human cells. A comparison was made between adduct formation in human cells and in purified DNA. The sequence-specific position and relative intensity of damage was similar in both systems for cisplatin, dichloro(ethylenediammine)platinum(II) (PtenCl2), and N-[3-N-(ethylenediamino)propyl]acridine-4-carboxamidedichloropl atinum(II) (4AcC3PtenCl2). However, no DNA damage could be detected in cells for trans-diamminedichloroplatinum(II) (transPt) or N-[3-N-(ethylenediamino)propyl]acridine-2-carboxamide-dichloroplat inum(II) (2AcC3PtenCl2) despite the ability of these latter analogues to damage purified DNA. Cisplatin, PtenCl2, and 4AcC3PtenCl2, which significantly damaged DNA inside cells, also show antitumor activity in mouse models. However, transPt and 2AcC3PtenCl2, which did not detectably damage DNA inside cells, did not show such antitumor activity. This correlation between intracellular DNA damaging ability and in vivo antitumor activity indicates the potential use of the human cells/Taq DNA polymerase/linear amplification technique as a convenient method for screening new cisplatin analogues for useful chemotherapeutic activity.
Murray
V
V
School of Biochemistry and Molecular Genetics, University of New South Wales, Kensington, Australia.
Motyka
H
H
England
P R
PR
Wickham
G
G
Lee
H H
HH
Denny
W A
WA
McFadyen
W D
WD
D000166:Acridines / Q000378:metabolism / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D002945:Cisplatin / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage / Q000187:drug effects; D004259:DNA-Directed DNA Polymerase; D005784:Gene Amplification; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009944:Organoplatinum Compounds / Q000378:metabolism / Q000494:pharmacology; D013054:Spectrophotometry, Atomic; D019914:Taq Polymerase
D000166:Acridines; D000970:Antineoplastic Agents; D009944:Organoplatinum Compounds; C002892:platinum ethylenediamine dichloride; C078353:N-(3-N-(ethylenediamino)propyl)acridine-4-carboxamide dichloroplatinum(II); D004247:DNA; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase; D002945:Cisplatin