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Transcriptional modulation of BCRP gene to reverse multidrug resistance by toremifene in breast adenocarcinoma cells

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Abstract

Breast cancer resistance protein (BCRP/ABCG2), an ATP-binding cassette half transporter, confers multidrug resistance (MDR) to a series of antitumor agents such as mitoxantrone, daunorubicin, SN-38, and topotecan, and often limits the efficacy of chemotherapy. Recent studies have indicated that a putative estrogen response element (ERE) is located in the promoter region of the BCRP gene. However, whether and how BCRP is regulated transcriptionally by toremifene (TOR) remains unknown. In the present study, two plasmid vectors have been designed to express the wild-type full-length BCRP cDNA enforced driven by its endogenous promoter containing a functional ERE and a constitutive cytomegalovirus (CMV) promoter as control, respectively, which were transfected into estrogen-responsive MCF-7 and estrogen-independent MDA-MB-231 human breast adenocarcinoma cell lines. We showed that toremifene alone significantly downregulated BCRP mRNA and protein levels in estrogen receptor α (ERα)-positive MCF-7 cells in a dose-dependent manner, and the inhibitory effect was partially reversed by estrone (E1). Furthermore, gel shift assays demonstrated that specific binding of ERα to the ERE in the BCRP promoter is essential for transcriptional inhibition of BCRP by toremifene. Interestingly, toremifene alone increased the cellular accumulation of mitoxantrone in BCRP-transfected cells, suggesting that TOR indeed inhibits BCRP-mediated drug efflux and overcome drug resistance. To the best of our knowledge, this is the first report describing a direct effect of toremifene on BCRP. Our results thus indicate that toremifene by itself downregulates BCRP expression to reverse BCRP-mediated atypical multidrug resistance via a novel transcriptionally mechanism, which might be involved in TOR–ER complexes binding to the ERE of BCRP promoter to repress transcription of BCRP gene.

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Acknowledgments

We thank Dr. Faliang Zhu (Department of Immunology, School of Medicine, Shandong University, China) for excellent technical assistance with flow cytometry. This research was supported by the National Natural Science Foundation of China (NSFC, No. 30300124) and the Specialized Research Fund for the Doctoral Program of Higher Education, People’s Republic of China (Project No. 20020422042).

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Authors and Affiliations

  1. Department of Pathology, School of Medicine, Shandong University, 44 Wenhuaxi Road, Jinan, 250012, People’s Republic of China

    Yuhua Zhang, Peng Gao, Xiaofang Zhang & Gengyin Zhou

  2. Department of Pathology, Beijing Tiantan Hospital, Capital Medical University, Beijing, 100050, People’s Republic of China

    Yuhua Zhang, Guang Li & Jin Yu

  3. Department of Pathology, 301 Hospital, Beijing, 100853, People’s Republic of China

    Huaiping Wang

  4. Department of Urology, Shandong Provincial Hospital, Jinan, People’s Republic of China

    Lijing Wei

  5. Department of Pathology, Shandong Cancer Hospital, Shandong Academy of Medical Science, 440 Jiyan Road, Jinan, Shandong, People’s Republic of China

    Yan Gao

  6. Department of Orthodontics, School of Stomatology, Shandong University, Jinan, 250012, People’s Republic of China

    Fulan Wei

  7. Department of Internal Medicine, Department of Pharmacology, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN, 37614, USA

    Deling Yin

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  1. Yuhua Zhang
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  11. Gengyin Zhou
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Correspondence to Gengyin Zhou.

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Zhang, Y., Wang, H., Wei, L. et al. Transcriptional modulation of BCRP gene to reverse multidrug resistance by toremifene in breast adenocarcinoma cells. Breast Cancer Res Treat 123, 679–689 (2010). https://doi.org/10.1007/s10549-009-0660-2

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  • DOI: https://doi.org/10.1007/s10549-009-0660-2

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