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

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...

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Veröffentlicht in:Clinical chemistry (Baltimore, Md.) Md.), 1998-03, Vol.44 (3), p.556-559
Hauptverfasser: Lambooy, Lambert H. J, Leegwater, Peter A. J, van den Heuvel, Lambert P, Bokkerink, Jos P, De Abreu, Ronney A
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Sprache:eng
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Zusammenfassung: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 the 14C/3H ratio as a measure for the methylation of newly formed DNA. The 14C/3H ratio was decreased by 17% with 1 mumol/L 6-MP; treatment with increasing concentrations of 6-MP up to 10 mumol/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.
ISSN:0009-9147
1530-8561
DOI:10.1093/clinchem/44.3.556