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Constitutive Overexpression of P-glycoprotein, Rather than Breast Cancer Resistance Protein or Organic Cation Transporter 1, Contributes to Acquisition of Imatinib-Resistance in K562 Cells

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Abstract

Purpose

The purpose of this study was to investigate the contribution of drug transporters in acquired imatinib-resistance. Specifically, we focused on the efflux transporters, P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), and an influx transporter, organic cation transporter 1 (OCT1).

Materials and methods

We established imatinib-resistant K562 cells (K562/IM). Real-time PCR or Western blot analyses were performed to examine the mRNA or protein levels. Alamar blue method was used in the cytotoxicity assay. The transport activities and intracellular imatinib levels were measured by flow cytometry and HPLC, respectively.

Results

K562/IM displayed a 47-fold increase in resistance to imatinib over the parent K562 cells. Both P-gp and BCRP were overexpressed in K562/IM relative to K562. Furthermore, the intracellular imatinib level was markedly reduced in K562/IM. Interestingly, cyclosporin A, a P-gp inhibitor, but not fumitremorgin C, a BCRP inhibitor, restored both imatinib-sensitivity and the intracellular imatinib level. By contrast, no significant difference in the expression and function of OCT1 was observed between K562/IM and K562.

Conclusions

P-gp, rather than BCRP or OCT1, is partially responsible for the development of imatinib-resistance due to constitutive and functional overexpression, leading to reduced intracellular accumulation of imatinib in K562/IM.

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Abbreviations

BCRP:

breast cancer resistance protein

IM:

imatinib

OCT1:

organic cation transporter 1

P-gp:

P-glycoprotein

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Acknowledgments

We thank Dr. Robert W. Robey and Dr. Susan E. Bate at National Cancer Institute (Betheda, MD) for providing FTC and PhA. We also acknowledge Novartis Pharma AG (Basel, Switzerland) for kindly providing imatinib and cyclosporin A.

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Authors and Affiliations

  1. Department of Pharmacy, Kumamoto University Hospital, 1-1-1 Honjo, Kumamoto, 860-8556, Japan

    Chie Hirayama, Hiroshi Watanabe, Reiko Nakashima, Takeru Nanbu, Akinobu Hamada & Hideyuki Saito

  2. Department of Molecular Cell Function, Graduate School of Medical and Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto, 862-0973, Japan

    Akihiko Kuniyasu & Hitoshi Nakayama

  3. Department of Hematology and Infectious Diseases, Kumamoto University Hospital, 1-1-1 Honjo, Kumamoto, 860-8556, Japan

    Tatsuya Kawaguchi

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  1. Chie Hirayama
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  2. Hiroshi Watanabe
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  9. Hideyuki Saito
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Correspondence to Hideyuki Saito.

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Hirayama, C., Watanabe, H., Nakashima, R. et al. Constitutive Overexpression of P-glycoprotein, Rather than Breast Cancer Resistance Protein or Organic Cation Transporter 1, Contributes to Acquisition of Imatinib-Resistance in K562 Cells. Pharm Res 25, 827–835 (2008). https://doi.org/10.1007/s11095-007-9376-3

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  • DOI: https://doi.org/10.1007/s11095-007-9376-3

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