Open AccessInhibition of DNA methylation in malignant MOLT F4 lymphoblasts by 6-mercaptopurine
Author(s) -
Lambert H.J. Lambooy,
Peter A. J. Leegwater,
Lambert P. van den Heuvel,
Jos P.M. Bökkerink,
Ronney A. De Abreu
Publication year - 1998
Publication title -
clinical chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.705
H-Index - 218
eISSN - 1530-8561
pISSN - 0009-9147
DOI - 10.1093/clinchem/44.3.556
Subject(s) - mercaptopurine , lymphoblast , methylation , dna methylation , dna , microbiology and biotechnology , chemistry , medicine , cancer research , biology , pharmacology , genetics , cell culture , gene , gene expression
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.
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