TY - JOUR
T1 - Cisplatin DNA adduct detection and depurination measured by 32P DNA radiolabeling and two-dimensional thin-layer chromatography
T2 - A time and concentration study
AU - Farah, Nazih
AU - Dresner, Harley S.
AU - Searles, Kenneth J.
AU - Winiarsky, Raz
AU - Moosikasuwan, Mark
AU - Cajigas, Antonio
AU - Hahm, Sae
AU - Steinberg, Jacob J.
PY - 2000
Y1 - 2000
N2 - Platinum-based chemotherapies cause the formation of DNA adducts and have profound effects on DNA. This study measured cis- diamminedichloroplatinum II (cisplatin) DNA adducts by 32P-radiolabeling DNA, enzymatically digesting radiolabeled DNA, separating the formed adducts on two-dimensional thin-layer chromatography, and quantitating the adducts with autoradiography and densitometry. HeLa DNA was incubated with cisplatin at varying concentrations (6.25-325 nM) and times (0 min to 72 hr). Cisplatin rapidly depurinated dGMP and dAMP (90%, 15-min incubation with 325 nM cisplatin). Partial depurination of dGMP (15%) and dAMP (25%) occurred with lower cisplatin concentrations at equal incubation times. A minimum of four new adducts, with relatively rapid migratory patterns, were detected at high cisplatin concentrations with short incubation times. These results indicate that the depurination of DNA correlates with DNA adduct formation and that the quantification of these adducts may be applicable to monitoring tumor and host cell response to cisplatin chemotherapy.
AB - Platinum-based chemotherapies cause the formation of DNA adducts and have profound effects on DNA. This study measured cis- diamminedichloroplatinum II (cisplatin) DNA adducts by 32P-radiolabeling DNA, enzymatically digesting radiolabeled DNA, separating the formed adducts on two-dimensional thin-layer chromatography, and quantitating the adducts with autoradiography and densitometry. HeLa DNA was incubated with cisplatin at varying concentrations (6.25-325 nM) and times (0 min to 72 hr). Cisplatin rapidly depurinated dGMP and dAMP (90%, 15-min incubation with 325 nM cisplatin). Partial depurination of dGMP (15%) and dAMP (25%) occurred with lower cisplatin concentrations at equal incubation times. A minimum of four new adducts, with relatively rapid migratory patterns, were detected at high cisplatin concentrations with short incubation times. These results indicate that the depurination of DNA correlates with DNA adduct formation and that the quantification of these adducts may be applicable to monitoring tumor and host cell response to cisplatin chemotherapy.
UR - http://www.scopus.com/inward/record.url?scp=0034037987&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034037987&partnerID=8YFLogxK
U2 - 10.3109/07357900009012174
DO - 10.3109/07357900009012174
M3 - Article
C2 - 10808367
AN - SCOPUS:0034037987
SN - 0735-7907
VL - 18
SP - 314
EP - 326
JO - Cancer Investigation
JF - Cancer Investigation
IS - 4
ER -