The Multidrug Resistance Protein MRP1 (ABCC1) can confer resistance to a variety of therapeutic drugs. In addition, MRP1/ABCC1 mediates cellular export of natural folates, such as folic acid and L-leucovorin. In this study we determined whether cellular folate status affected the functional activity of MRP1/ABCC1 mediated efflux of an established substrate, the anthracycline daunorubicin (DNR). As a model system we used the human ovarian carcinoma cell line 2008wt, and its MRP1/ABCC1 transfected subline 2008/MRP1. Both types of these moderate- and high-MRP1/ABCC1 expressing cells displayed efflux of DNR when maintained in standard culture media (2.3μM folic acid). The initial total cellular DNR efflux rate in 2008/MRP1 cells was ∼2-fold higher compared to 2008wt cells. This efflux consisted of MRP1/ABCC1 mediated transport, possibly non-MRP1 mediated transport, as well as passive diffusion. Benzbromarone, a specific MRP1 inhibitor, decreased the initial efflux rate in 2008/MRP1 cells (4-fold) and in 2008wt cells (2-fold). When 2008/MRP1 cells were challenged for 2 days in folate-free medium, total cellular DNR efflux was decreased to 43% of the initial efflux rate under folate-rich conditions. In 2008wt cells DNR efflux was decreased to 84% of the folate-rich conditions. Benzbromarone did not inhibit DNR efflux after the folate-free period in both cell lines. Repletion of folate by a 2-24hr exposure to 2.5μM L-leucovorin or folic acid resulted in a complete restoration of DNR efflux. In contrast, expression of MRP1/ABCC1 protein was not changed significantly during the folate-free period or the repletion-period, nor were cellular ATP or ADP pools. In conclusion, this study demonstrates that the cellular folate status can influence the transport activity of MRP1/ABCC1. These results have potentially important implications in the understanding of the (patho-)physiological roles of MRP1/ABCC1, and possibly other ABC transporter proteins in cellular folate homeostasis and drug resistance.

ABCC1, Daunorubicin, DHFR, Dihydrofolate reductase, DNR, Folate, Folylpolyglutamyl synthetase, FPGS, HF, High-folate, L-leucovorin, LF, Low-folate, LV, Methotrexate, MRP1, MTX, Multidrug Resistance Protein, Reduced folate carrier, RFC, Thymidylate synthase, TS,
Biochemical Pharmacology
Department of Pediatrics

Hooijberg, J.H, Jansen, G, Assaraf, Y.G, Kathmann, I, Pieters, R, Laan, A.C, … Peters, G.J. (2004). Folate concentration dependent transport activity of the Multidrug Resistance Protein 1 (ABCC1). Biochemical Pharmacology, 67(8), 1541–1548. doi:10.1016/j.bcp.2003.12.022