Biochemical modulation of Ara-C effects by amidox, an inhibitor of ribonucleotide reductase in HL-60 promyelocytic human leukemia cells.
Amidox, a new polyhydroxy-substituted benzoic acid derivative, is a potent inhibitor of the enzyme ribonucleotide reductase (RR), which catalyses the de novo synthesis of DNA. RR is considered to be an excellent target for anti cancer chemotherapy. We investigated the biochemical and antineoplastic effects of amidox as a single agent and in combination with Ara-C in human HL-60 promyelocytic leukemia cells. Amidox inhibited the growth of HL-60 cells in a growth inhibition assay with an IC50 of 25 microM. In a soft agar colony forming assay, amidox yielded a 50% inhibition of colony formation at 13 microM. We also investigated the effects of amidox treatment on the formation of deoxynucleosidetriphosphates. Amidox (50 and 75 microM for 24 hours) could significantly decrease intracellular concentrations of dCTP, dATP and dGTP pools, whereas dTTP levels increased. We then tested the combination effects of amidox with Ara-C; this combination yielded additive cytotoxic effects both in growth inhibition and in soft agar colony formation assays. This effect was due to the increased formation of Ara-CTP, the active metabolite of Ara-C, after preincubation with amidox. Preincubation of HL-60 cells with 75 and 100 microM amidox for 24 hours caused an increase in the intracellular Ara-CTP concentrations by 576% and 1143%, respectively. Therefore amidox might offer an additional option for the treatment of leukemia and thus be further investigated in in vivo studies as a single agent and in combination with Ara-C.
Höchtl
Thomas
T
Clinical Institute for Medical and Chemical Laboratory Diagnostics, School of Medicine, University of Vienna, General Hospital of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria.
Horvath
Zsuzsanna
Z
Bauer
Wolfgang
W
Karl
Doris
D
Saiko
Philipp
P
Elford
Howard L
HL
Fritzer-Szekeres
Monika
M
Szekeres
Thomas
T
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D001085:Arabinofuranosylcytosine Triphosphate / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D003114:Colony-Forming Units Assay; D003561:Cytarabine / Q000494:pharmacology*; D003854:Deoxyribonucleotides / Q000378:metabolism; D006131:Growth Inhibitors / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D010091:Oximes / Q000494:pharmacology*; D012264:Ribonucleotide Reductases / Q000037:antagonists & inhibitors*
D000964:Antimetabolites, Antineoplastic; D003854:Deoxyribonucleotides; D006131:Growth Inhibitors; D010091:Oximes; D003561:Cytarabine; D001085:Arabinofuranosylcytosine Triphosphate; C070491:Amidox; D012264:Ribonucleotide Reductases