Clinical Chemistry
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Clinical Chemistry 44: 556-559, 1998;
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(Clinical Chemistry. 1998;44:556-559.)
© 1998 American Association for Clinical Chemistry, Inc.

Drug Monitoring and Toxicology

Inhibition of DNA methylation in malignant MOLT F4 lymphoblasts by 6-mercaptopurine

Lambert H. J. Lambooy, Peter A. J. Leegwater, Lambert P. van den Heuvel, Jos P. Bökkerink, and Ronney A. De Abreua

a Author for correspondence. Fax (+)31-24-3616428; e-mail r.deabreu{at}ckslkn.azn.nl.

Treatment of MOLT F4 lymphoblasts with 6-mercaptopurine (6-MP) resulted in a decrease of ATP and a depletion of S-adenosylmethionine (AdoMet). To investigate whether this might affect the methylation of DNA, we treated MOLT F4 lymphoblasts with increasing concentrations of 6-MP, followed by labeling with [methyl-14C]methionine and [methyl-3H]thymidine. After DNA isolation, we measured the incorporated radioactivity and determined the14C/3H ratio as a measure for the methylation of newly formed DNA. The 14C/3H ratio was decreased by 17% with 1 µmol/L 6-MP; treatment with increasing concentrations of 6-MP up to 10 µmol/L showed a further decrease to 70%, in comparison with untreated cells. To demonstrate that the methylation of deoxycytidine residues in DNA was reduced, we quantified hydrolyzed DNA by HPLC. The 14C/3H ratio showed a decrease with increasing 6-MP concentrations, indicating that treatment with 6-MP resulted in hypomethylation of DNA.






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Copyright © 1998 by the American Association for Clinical Chemistry.