T. Dervieux et al., Differing contribution of thiopurine methyltransferase to mercaptopurine versus thioguanine effects in human leukemic cells, CANCER RES, 61(15), 2001, pp. 5810-5816
Thioguanine and mercaptopurine are prodrugs requiring conversion into thiop
urine nucleotides to exert cytotoxicity. Thiopurine S-methyltransferase (TP
MT), an enzyme subject to genetic polymorphism, catabolizes thiopurines int
o inactive methylated bases, but also produces methylthioguanine nucleotide
s and methylmercaptopurine nucleotides from thioguanine and mercaptopurine
nucleotides, respectively. To study the effect of TPMT on activation versus
inactivation of mercaptopurine and thioguanine, we used a retroviral gene
transfer technique to develop human CCRF-CEM cell lines that did (TPMT+) an
d did not (MOCK) overexpress TPMT. After transduction, TPMT activities were
14-fold higher in the TPMT+ versus the MOCK cell lines (P < 0.001). TPMTcells were less sensitive to thioguanine than MOCK cells (IC50 = 1.10 +/- 0
.12 muM versus 0.55 +/- 0.19 muM; P = 0.02); in contrast, TPMT+ cells were
more sensitive to mercaptopurine than MOCK cells (IC50 = 0.52 +/- 0.20 muM
versus 1.50 +/- 0.23 muM; P < 0.01). The lower sensitivity of TPMT+ versus
MOCK cells to thioguanine was associated with lower thioguanine nucleotide
concentrations (917 +/- 282 versus 1515 +/- 183 pmol/5 X 10(6) cells; P = 0
.01), higher methylthioguanine nucleotide concentrations (252 +/- 34 versus
27 +/- 10 pmol/5 X 106 cells; P = 0.01), less inhibition of de novo purine
synthesis (13 versus 95%; P < 0.01), and lower deoxythioguanosine incorpor
ation into DNA (2.0 +/- 0.6% versus 7.2 +/- 2.0%; P < 0.001). The higher se
nsitivity of TPMT+ cells to mercaptopurine was associated with higher conce
ntrations of methylmercaptopurine nucleotide (2601 +/- 1055 versus 174 +/-
77 pmol/5 X 106 Cells; P = 0.01) and greater inhibition of de novo purine s
ynthesis (> 99% versus 74%; P < 0.01) compared with MOCK cells. We conclude
that methylation of mercaptopurine contributes to the antiproliferative pr
operties of the drug, probably through inhibition of de novo purine synthes
is by methylmercaptopurine nucleotides, whereas thioguanine is inactivated
primarily by TPMT.