Variable expression of RFC1 in human leukemia cell lines resistant to antifolates.
The resistance to folate-based antifolates is associated with impaired function of the reduced folate carrier (RFC), one of the major routes of folate transport into cancer cells. To clarify the importance of RFC functions in the antifolate resistance, we have examined the expression of RFC1 and its phenotype as a folate transporter in human leukemia cell lines resistant to various antifolates. MOLT-3 cells resistant to ZD9331 (a thymidylate synthase (TS) inhibitor that utilizes the RFC for cell entry) (MOLT-3/ZD9331) showed decreased expression of RFC1 concomitant with diminished cellular uptake of [3H]methotrexate (MTX). K562 cells resistant to raltitrexed (ZD1694, another TS inhibitor that utilizes the RFC for cell entry) (K562/ ZD1694 x C) scarcely expressed RFC1, which is in accordance with the impaired uptake of folate analogs and the high degree of resistance to ZD1694 and MTX. On the other hand, no apparent decrease of RFCI1 expression was found in transport-deficient MTX-resistant MOLT-3 cells (MOLT-3/MTX10000) though its phenotype showed defective transport of MTX or ZD1694. In these cell lines with impaired RFC function, [3H]leucovorin (LV) uptake was only moderately decreased as compared to [3H]MTX or [3H]ZD1694 uptake. These cells grew with a minimal retardation in folate-free medium supplemented with 10 nM LV, suggesting that these cell lines with impaired RFC function had enough folate transporters to transport LV. In contrast to downregulation of RFC, the much greater uptake of [3H]MTX was observed in the MOLT-3/trimetrexate (TMQ)800-MTX10000 in parallel with increased RFC1 expression. These cell lines with the altered expression of RFC1 may serve as models useful for investigating the regulation of RFC1 expression and for understanding the molecular mechanism(s) behind the transport-mediated antifolate resistance.
Kobayashi
H
H
Department of Laboratory Medicine, National Defense Medical College, Tokorozawa, Saitama, Japan. hkobayas@is.icc.u-tokai.ac.jp
Takemura
Y
Y
Ohnuma
T
T
D000964:Antimetabolites, Antineoplastic / Q000493:pharmacokinetics; D001692:Biological Transport; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000378:metabolism; D019008:Drug Resistance, Neoplasm; D005492:Folic Acid / Q000378:metabolism / Q000494:pharmacology; D005493:Folic Acid Antagonists / Q000493:pharmacokinetics / Q000494:pharmacology*; D006801:Humans; D002955:Leucovorin / Q000493:pharmacokinetics; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000188:drug therapy* / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000378:metabolism; D026901:Membrane Transport Proteins; D008727:Methotrexate / Q000493:pharmacokinetics; D010641:Phenotype; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy* / Q000378:metabolism*; D011799:Quinazolines / Q000493:pharmacokinetics; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism; D013876:Thiophenes / Q000493:pharmacokinetics; D014407:Tumor Cells, Cultured
D000964:Antimetabolites, Antineoplastic; D002352:Carrier Proteins; D005493:Folic Acid Antagonists; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D011799:Quinazolines; C120043:SLC19A2 protein, human; D013876:Thiophenes; D005492:Folic Acid; D013762:Tetrahydrofolate Dehydrogenase; C068874:raltitrexed; D002955:Leucovorin; D008727:Methotrexate