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The Identification of Two Germ-line Mutations in the Human Breast Cancer Resistance Protein Gene that Result in the Expression of a Low/Non-functional Protein

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Abstract

Purpose

We examined the effects of the nine nonsynonymous germ-line mutations/SNPs in the breast cancer resistance protein (BCRP/ABCG2) gene on the expression and function of the protein.

Materials and Methods

We generated cDNAs for each of these mutants (G151T, C458T, C496G, A616C, T623C, T742C, T1291C, A1768T, and G1858A BCRP) and compared the effects of their exogenous expression in PA317 cells with a wild-type control.

Results

PA/F208S cells (T623C BCRP-transfectants) expressed marginal levels of a BCRP protein species (65kDa), which is slightly smaller than wild-type (70kDa), but this mutant did not appear on the cell surface or confer drug resistance. PA/F431L cells (T1291C BCRP-transfectants) were found to express both 70kDa and 65kDa BCRP protein products. In addition, although PA/F431L cells expressed 70kDa BCRP at comparable levels to PA/WT cells, they showed only marginal resistance to SN-38. PA/T153M cells (C458T BCRP-transfectants) and PA/D620N cells (G1858A BCRP-transfectants) expressed lower amounts of BCRP and showed lower levels of resistance to SN-38 compared with PA/WT cells.

Conclusions

We have shown that T623C BCRP encodes a non-functional BCRP and that T1291C BCRP encodes a low-functional BCRP. Hence, these mutations may affect the pharmacokinetics of BCRP substrates in patients harboring these alleles.

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Abbreviations

ABC transporter:

ATP-binding cassette transmembrane transporter

BCRP:

breast cancer resistance protein

DHFR:

dihydrofolate reductase

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

IRES:

internal ribosome entry site

SNPs:

single nucleotide polymorphisms

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Acknowledgments

This study was supported by the Ministry of Education, Culture, Sports, Science and Technology, and the Ministry of Health, Labor and Welfare, Japan.

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Authors and Affiliations

  1. Department of Chemotherapy, Kyoritsu University of Pharmacy, 1-5-30 Shibakoen, Minato-ku, Tokyo, 105-8512, Japan

    Sho Yoshioka, Kazuhiro Katayama, Chikako Okawa, Sachiko Takahashi, Junko Mitsuhashi & Yoshikazu Sugimoto

  2. Division of Gene Therapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Tokyo, Japan

    Satomi Tsukahara, Junko Mitsuhashi & Yoshikazu Sugimoto

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  1. Sho Yoshioka
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  2. Kazuhiro Katayama
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Correspondence to Yoshikazu Sugimoto.

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Yoshioka, S., Katayama, K., Okawa, C. et al. The Identification of Two Germ-line Mutations in the Human Breast Cancer Resistance Protein Gene that Result in the Expression of a Low/Non-functional Protein. Pharm Res 24, 1108–1117 (2007). https://doi.org/10.1007/s11095-007-9235-2

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  • DOI: https://doi.org/10.1007/s11095-007-9235-2

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