Overexpression of wild-type breast Cancer resistance protein mediates methotrexate resistance

• Published in: Cancer research (Chicago, Ill.) Ill.), 2002-09, Vol.62 (17), p.5035-5040
• Main Authors: VOLK, Erin L, FARLEY, Kate M, YAN WU, FEI LI, ROBEY, Robert W, SCHNEIDER, Erasmus
• Format: Article
• Language: English
• Subjects: Antimetabolites, Antineoplastic - pharmacology; Antineoplastic agents; ATP Binding Cassette Transporter, Sub-Family G, Member 2; ATP-Binding Cassette Transporters - antagonists & inhibitors; ATP-Binding Cassette Transporters - biosynthesis; ATP-Binding Cassette Transporters - genetics; Biological and medical sciences; Breast Neoplasms - drug therapy; Breast Neoplasms - genetics; Breast Neoplasms - metabolism; Chemotherapy; Drug Resistance, Neoplasm; Gene Amplification; Humans; Medical sciences; Methotrexate - pharmacokinetics; Methotrexate - pharmacology; Neoplasm Proteins; Pharmacology. Drug treatments; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Substrate Specificity; Tumor Cells, Cultured
• ISSN: 0008-5472; 1538-7445

Previously, we have reported that a multidrug-resistant, mitoxantrone (MX)-selected cell line, MCF7/MX, is highly cross-resistant to the antifolate methotrexate (MTX), because of enhanced ATP-dependent drug efflux (E. L. Volk et al., Cancer Res., 60: 3514-3521, 2000). These cells overexpress the breast cancer resistance protein (BCRP), and resistance to MTX as well as to MX was reversible by the BCRP inhibitor, GF120918. These data indicated that BCRP causes the multidrug-resistance phenotype. To further examine the role of this transporter in MTX resistance, and in particular the role of amino acid 482, we analyzed a number of BCRP-overexpressing cell lines. MTX resistance correlated with BCRP expression in all of the cell lines expressing the wild-type transporter, which contains an Arg at position 482. In contrast, little or no cross-resistance was found in the MCF7/AdVp1000 and S1-M1-3.2 and S1-M1-80 cell lines, which contain acquired mutations at this position, R482T and R482G, respectively. Concomitantly, the greatest reduction in MTX accumulation was observed in the MCF7/MX cells (BCRP(Arg)) as compared with cells expressing the Thr and Gly BCRP variants. Furthermore, the reduction in drug accumulation was sensitive to BCRP inhibition by GF120918. In conclusion, we have demonstrated a novel role for BCRP as a mediator of MTX resistance and have provided further evidence for the importance of amino acid 482 in substrate specificity.