Molecular mechanisms underlying the enhanced sensitivity of thiopurine-resistant T-lymphoblastic cell lines to methyl mercaptopurineriboside.
Methylmercaptopurine riboside (meMPR), a cellular metabolite of 6-mercaptopurine (6-MP), is a potent inhibitor of de novo purine synthesis (DNPS). Human MOLT4 T-lymphoblastic leukaemia cells that have acquired resistance to 6-MP or 6-thioguanine (6-TG) as a consequence of defective transport exhibit enhanced sensitivity to meMPR. HPLC-based analysis of the transport of meMPR revealed normal uptake of this compound by our thiopurine-resistant cell sublines, suggesting a route of transport distinct from that for 6-MP and 6-TG. Studies on the wild-type parental leukemic cells showed that adenosine, dipyridamole and nitrobenzylthioinosine inhibit uptake of meMPR to a significant extent, whereas Na+ ions have no influence on this process. Transfection of these leukemic cells with small interference RNA molecules targeting the gene encoding the first member of the family of equiliberative nucleoside transporters (ENT1) strongly reduced the initial rate of meMPR transport. Our resistant cell lines exhibited 30-52% reductions (p < 0.005) in their levels of mRNA encoding several proteins involved in de novo purine synthesis, i.e., aminoimidazole carboxamide ribonucleotide formyltransferase, glycinamide ribonucleotide transformylase and guanine monophosphate synthetase. Consequently, the rate of de novo purine synthesis in these resistant sublines was decreased by 50%. Furthermore, the levels of ribonucleoside triphosphates in these cells were significantly lower than in the non-resistant parental cells. In combination, a reduced rate of de novo purine synthesis together with low levels of ribonucleoside triphosphates can explain the enhanced sensitivity of our thiopurine-resistant cell lines to meMPR. In this manner, meMPR bypasses the mechanisms of resistance to thiopurines and is even more cytotoxic towards resistant than towards wild-type cells.
Fotoohi
Alan Kambiz
AK
Department of Oncology and Pathology, Cancer Center Karolinska (CCK), Karolinska Hospital and Institute, 171 76 Stockholm, Sweden.
Wrabel
Anna
A
Moshfegh
Ali
A
Peterson
Curt
C
Albertioni
Freidoun
F
D000248:Adenosine Kinase / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D015536:Down-Regulation / Q000187:drug effects; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D033721:Equilibrative Nucleoside Transporter 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing; D006801:Humans; D020128:Inhibitory Concentration 50; D015122:Mercaptopurine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods; D011687:Purines / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D012265:Ribonucleotides / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D012964:Sodium / Q000502:physiology; D013601:T-Lymphocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D013866:Thioguanine / Q000494:pharmacology
D033721:Equilibrative Nucleoside Transporter 1; D011687:Purines; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012265:Ribonucleotides; C010240:6-methylthiopurine; D012964:Sodium; D015122:Mercaptopurine; D000248:Adenosine Kinase; D013866:Thioguanine