Deoxyribonucleoside triphosphates in human cells: changes in disease and following exposure to drugs.
Deoxyribonucleic acid synthesis requires adequate cellular concentrations of the four deoxyribonucleoside triphosphates. Using a sensitive enzymic assay, we have measured the concentrations (pools) of these compounds in human bone marrow cells and in lymphocytes. The mean concentrations (pmol/10(6) cells) in normal human bone marrow cells were: deoxyadenosine triphosphate (dATP) 1.5; deoxyguanosine triphosphate (dGTP) 0.4; thymidine triphosphate (dTTP) 1.4 and deoxycytidine triphosphate (dCTP) 0.6; and in normal phytohaemagglutinin (PHA)-stimulated lymphocytes (72 h cultures); dATP 3.7; dGTP 1.9; dTTP 9.4 and dCTP 2.9. The deoxyribonucleoside triphosphate concentrations were increased approximately threefold in the nucleated marrow cells from patients with leukaemia and myeloproliferative diseases. PHA-stimulation of lymphocytes caused a marked increase of the deoxyribonucleoside triphosphate concentrations, particularly of dTTP, between 24 and 48 h of culture. In PHA-stimulated lymphocytes, the antifolate drugs methotrexate, pyrimethamine and trimethoprim, all produced a fall in dTTP and a rise in dATP concentrations within 1 h. These effects could be reversed by folinic acid. 5-Fluorouracil caused a fall in dTTP and in dCTP but no consistent changes in dATP; hydroxyurea caused a fall in dATP with a rise in dTTP. BCNU caused a significant fall in dATP and dCTP. Dibutyryl cyclic 3', 5' adenosine monophosphate and theophylline had no consistent effect on the deoxyribonucleoside triphosphate concentrations. 6-Mercaptopurine caused a fall in dATP and dGTP, the fall in dATP being marked after 4 h incubation. It is concluded that measurement of the deoxyribonucleoside triphosphates in human cells provides a new method of studying DNA synthesis in human disease states and of analysing the action of antimetabolite drugs on normal and diseased cells.
D000255:Adenosine Triphosphate / Q000032:analysis; D000963:Antimetabolites / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology; D001853:Bone Marrow / Q000032:analysis*; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D003597:Cytosine Nucleotides / Q000032:analysis; D004247:DNA / Q000096:biosynthesis; D004273:DNA, Neoplasm / Q000096:biosynthesis; D003854:Deoxyribonucleotides / Q000032:analysis*; D006160:Guanosine Triphosphate / Q000032:analysis; D007938:Leukemia / Q000032:analysis; D008214:Lymphocytes / Q000032:analysis*; D009196:Myeloproliferative Disorders / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism; D013942:Thymine Nucleotides / Q000032:analysis
D000963:Antimetabolites; D000970:Antineoplastic Agents; D003597:Cytosine Nucleotides; D004273:DNA, Neoplasm; D003854:Deoxyribonucleotides; D013942:Thymine Nucleotides; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D004247:DNA; D013936:Thymidine