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miR-218 targets survivin and regulates resistance to chemotherapeutics in breast cancer

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Abstract

Multidrug resistance (MDR) remains one of the most significant obstacles in breast cancer treatment, and this process often involves dysregulation of a great number of microRNAs (miRNAs). Some miRNAs are indicators of drug resistance and confer resistance to chemotherapeutic drugs, although our understanding of this complex process is still incomplete. We have used a combination of miRNA profiling and real-time PCR in two drug-resistant derivatives of MCF-7 and Cal51 cells. Experimental modulation of miR expression has been obtained by retroviral transfection. Taxol and doxorubicin IC50 values were obtained by short-term drug sensitivity assays. Apoptosis was determined by flow cytometry after annexin V staining, by caspase 3/7 and caspase 9 activity assays and the levels of apoptosis-related proteins bcl-2 and bax by real-time PCR and Western blot. miR target was studied using transient transfection of luciferase constructs with the 3′ untranslated regions (UTR) of target mRNAs. Small interfering RNA-mediated genetic knock-down was performed in MDR cells and its modulatory effect on apoptosis examined. The effect of miRNA on tumorigenicity and tumor drug response was studied in mouse xenografts. miRNA profiling of two drug-resistant breast cancer cell models indicated that miR-218 was down-regulated in both MCF-7/A02 and CALDOX cells. Ectopic expression of miR-218 resensitized both drug-resistant cell lines to doxorubicin and taxol due to an increase in apoptosis. miR-218 binds survivin (BIRC5) mRNA 3′-UTR and down-regulated reporter luciferase activity. Experimental down-regulation of survivin by RNA interference in drug-resistant cells did mimic the sensitization observed when miRNA-218 was up-regulated. In addition, resensitization to taxol was also observed in mouse tumor xenografts from cells over-expressing miR-218. miR-218 is involved in the development of MDR in breast cancer cells via targeting survivin and leading to evasion of apoptosis. Targeting miR-218 and survivin may thus provide a potential strategy for reversing drug resistance in breast cancer.

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Acknowledgments

We thank Prof. Dongsheng Xiong (Institute of Hematology, PUMC, Tianjin, China) for a generous gift of human breast cancer MCF-7 and MCF-7/A02 cells. This work was supported by the Chinese National Natural Sciences Foundation (81402480 to YH), the Science and Technology Foundation of Tianjin Municipal Health Bureau (2014KZ078 to YH) and the China Scholarship Council (to YH) for visiting Imperial College London.

Conflict of interest

The authors declared no conflict of interest.

Ethical approval

All procedures performed in studies involving animals performed at the National Institutes of Health (Tianjin Cancer Hospital) in accordance with guidelines under the Institutional Animal Care and Use Committee (IACUC) and approved by the Committee on the Ethics of Animal Experiments of the Tianjin Cancer Hospital. All applicable national and institutional guidelines for the care and use of animals were followed.

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    Authors and Affiliations

    1. The 3rd Department of Breast Cancer, China Tianjin Breast Cancer Prevention, Treatment and Research Center, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Huan-Hu-Xi Road, Ti-Yuan-Bei, He xi District, Tianjin, People’s Republic of China

      Yunhui Hu

    2. State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, People’s Republic of China

      Kun Xu

    3. Division of Cancer, Cancer Research Center, Faculty of Medicine, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK

      Ernesto Yagüe

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    Correspondence to Yunhui Hu or Ernesto Yagüe.

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    Yunhui Hu, Kun Xu, and Ernesto Yagüe contributed equally to this study.

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    Hu, Y., Xu, K. & Yagüe, E. miR-218 targets survivin and regulates resistance to chemotherapeutics in breast cancer. Breast Cancer Res Treat 151, 269–280 (2015). https://doi.org/10.1007/s10549-015-3372-9

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