Abstract
ATP-binding cassette transporter ABCG2 [breast cancer resistance protein (BCRP)] is a member of the ABC transporter superfamily that actively extrudes xenotoxins from cells and is a major determinant of the bioavailability of many compounds. ABCG2 expression is strongly induced during lactation in the mammary gland and is related to the active secretion of drugs into the milk. The presence of drug residues and environmental pollutants in milk is an outstanding problem for human milk consumption and milk industrial processes, involving important risks to public health and the dairy industry. In cows, a single nucleotide polymorphism (SNP) in this protein has been described previously (Tyr581) and is associated with higher fat and protein percentages and lower milk yield. However, whether this amino acid substitution affects ABCG2-mediated drug transport in cows, including milk secretion, required further exploration. We cloned the two variants of bovine ABCG2 and evaluated the effect of this SNP on mitoxantrone accumulation assays performed in ovine primary fibroblasts transiently expressing either of the variants. It is interesting to note that statistically significant differences in activity between both variants were observed, and the Ser581 variant was related with an increased efflux activity. In addition, we demonstrated that genistein is a very good inhibitor of bovine ABCG2 and identified new inhibitors of the transporter, such as the macrocyclic lactones, ivermectin, and selamectin. Moreover, the inhibitory effect of these compounds on human and murine ABCG2 homologs was confirmed using transduced Marbin-Dabin canine kidney II cells. These findings may have important implications regarding the presence of drug residues in milk and drug interactions affecting the pharmacological behavior of ABCG2 substrates.
Footnotes
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This work was supported by Ministerio de Ciencia y Tecnología (Spain) [Grant BIO2006-00430]; a Ramón y Cajal grant from the Ministerio de Ciencia y Tecnología (Spain) [Grant RyC-2006-001896]; and University of Leon (Spain) [Grant ULE2006-4].
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G.M. and M.M.M. contributed equally to this work.
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Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
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doi:10.1124/dmd.108.022715.
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ABBREVIATIONS: ABC, ATP-binding cassette; BCRP, breast cancer resistance protein; SNP, single nucleotide polymorphism; MXR, mitoxantrone; PCR, polymerase chain reaction; IRES, internal ribosome entry site; GFP, green fluorescent protein; SBFF1, Scottish Black face fibroblasts; MDCKII, Marbin-Dabin canine kidney II; PBS, phosphate-buffered saline; MF, median of fluorescence.
- Received June 5, 2008.
- Accepted September 24, 2008.
- The American Society for Pharmacology and Experimental Therapeutics
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