The kinase inhibitor O6-cyclohexylmethylguanine (NU2058) potentiates the cytotoxicity of cisplatin by mechanisms that are independent of its effect upon CDK2.
O(6)-Cyclohexylmethylguanine (NU2058) was developed as an inhibitor of CDK2 and was previously shown to potentiate cisplatin cytotoxicity in vitro. The aim of this study was to investigate the mechanism of cisplatin potentiation by NU2058. SQ20b, head and neck cancer cells were treated for 2h with NU2058 (100 microM) and then for a further 2h with cisplatin and NU2058. NU2058 increased cisplatin cytotoxicity, by clonogenic assay, with a dose modification factor (DMF) of 3.1. NU2058 increased total intracellular platinum levels 1.5-fold, and platinum-DNA adduct levels twofold. Furthermore, the cisplatin-DNA adducts formed were more toxic in the presence of NU2058. To investigate whether the effects of NU2058 on cisplatin adduct levels and toxicity were dependent on CDK2 activity, additional CDK2 inhibitors were tested. NU6230 (CDK2 IC(50) 18 microM) was equipotent to NU2058 (CDK2 IC(50) 17 microM) as a CDK2 inhibitor in cell-free and cell-based assays, yet did not potentiate cisplatin cytotoxicity. Furthermore, NU6102 was >1000-fold more potent than NU2058 as a CDK2 inhibitor (CDK2 IC(50) 5 nM) yet was no more active than NU2058 in potentiating cisplatin. NU2058 also potentiated melphalan (DMF 2.3), and monohydroxymelphalan (1.7), but not temozolomide or ionising radiation. Whilst NU2058 increased melphalan cytotoxicity, it did not increase melphalan-DNA adduct formation. These studies demonstrate that NU2058 alters the transport of cisplatin, causing more Pt-DNA adducts, as well as sensitizing cells to cisplatin- and melphalan-induced DNA damage. However, the effects of NU2058 are independent of CDK2 inhibition.
Northern Institute for Cancer Research, Newcastle University, Paul O'Gorman Building, Newcastle upon Tyne NE2 4HH, United Kingdom.
Luke R E
LR
Harrison
Christopher J
CJ
Ottley
D Graham
DG
Pearson
Céline
C
Roche
Stephen R
SR
Wedge
M Eileen
ME
Dolan
David R
DR
Newell
Michael J
MJ
Tilby
D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects*; D002945:Cisplatin / Q000378:metabolism / Q000494:pharmacology*; D051357:Cyclin-Dependent Kinase 2 / Q000037:antagonists & inhibitors*; D018736:DNA Adducts / Q000378:metabolism; D004357:Drug Synergism; D006147:Guanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D014410:Tumor Stem Cell Assay
D000970:Antineoplastic Agents; D018736:DNA Adducts; C466913:NU2058; D047428:Protein Kinase Inhibitors; D006147:Guanine; D051357:Cyclin-Dependent Kinase 2; D002945:Cisplatin