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Exosomal miR-221/222 enhances tamoxifen resistance in recipient ER-positive breast cancer cells

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Abstract

Recent studies have demonstrated that specific miRNAs, such as miR-221/222, may be responsible for tamoxifen resistance in breast cancer. Secreted miRNAs enclosed in exosomes can act as intercellular bio-messengers. Our objective is to investigate the role of secreted miR-221/222 in tamoxifen resistance of ER-positive breast cancer cells. Transmission electron microscopy analysis and nanoparticle tracking analysis were performed to determine the exosomes difference between MCF-7TamR (tamoxifen resistant) and MCF-7wt (tamoxifen sensitive) cells. PKH67 fluorescent labeling assay was used to detect exosomes derived from MCF-7TamR cells entering into MCF-7wt cells. The potential function of exosomes on tamoxifen resistance transmission was analyzed with cell viability, apoptosis ,and colony formation. MiRNA microarrays and qPCR were used to detect and compare the miRNAs expression levels in the two cells and exosomes. As the targets of miR-221/222, p27 and ERα were analyzed with western blot and qPCR. Compared with the MCF-7wt exosomes, there were significant differences in the concentration and size distribution of MCF-7TamR exosomes. MCF-7wt cells had an increased amount of exosomal RNA and proteins compared with MCF-7TamR cells. MCF-7TamR exosomes could enter into MCF-7wt cells, and then released miR-221/222. And the elevated miR-221/222 effectively reduced the target genes expression of P27 and ERα, which enhanced tamoxifen resistance in recipient cells. Our results are the first to show that secreted miR-221/222 serves as signaling molecules to mediate communication of tamoxifen resistance.

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Abbreviations

MCF-7TamR :

Tamoxifen-resistant MCF-7

MCF-7wt :

Tamoxifen-sensitive MCF-7

TAM:

Tumor-associated macrophage

ERα:

Estrogen receptor α

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Acknowledgments

We thank Shuai Zhang for his excellent technical support of nanoparticle tracking analysis. This study was supported by National Natural Science Foundation of China (Nos. 81202091, 81372390 and 81102006).

Conflict of interest

The authors declare no conflict of interest.

Author information

Authors and Affiliations

  1. State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, The Fourth Military Medical University, Xi’an, 710032, Shaanxi, China

    Yifang Wei, Xiaofeng Lai, Yongzheng Ma, Libo Yao & Jian Zhang

  2. Cadet Brigade of the Fourth Military Medical University, The Fourth Military Medical University, Xi’an, 710032, Shaanxi, China

    Shentong Yu & Huichen Li

  3. Department of Pharmacy, Xijing Hospital, The Fourth Military Medical University, Xi’an, 710032, Shaanxi, China

    Suning Chen

  4. The State Key Discipline of Cell Biology, Department of Oncology, Xijing Hospital, The Fourth Military Medical University, Xi’an, 710032, Shaanxi, China

    Yuan Zhang

  5. Department of the Key Laboratory, The First Affiliated Hospital, Xi’an Medical College, Xi’an, 710077, Shaanxi, China

    Xingmei Zhu

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  1. Yifang Wei
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  2. Xiaofeng Lai
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  4. Suning Chen
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  6. Yuan Zhang
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  7. Huichen Li
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  8. Xingmei Zhu
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  9. Libo Yao
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  10. Jian Zhang
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Corresponding authors

Correspondence to Libo Yao or Jian Zhang.

Additional information

Yifang Wei and Xiaofeng Lai contributed equally to this study.

Electronic supplementary material

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10549_2014_3037_MOESM1_ESM.tif

Fig. S1 Quantification of mean diameters of the exosomes isolated from MCF-7wt and MCF-7TamR cells. Diameters of exosomes were manually measured in each of five random microscopic fields (TIFF 2071 kb)

Fig. S2 Quantification of particles were analyzed by nanoparticle tracking analysis (NTA) (TIFF 422 kb)

10549_2014_3037_MOESM3_ESM.tif

Fig. S3 T47DTamR exosomes deliver miR-221/222 into recipient T47Dwt cells (a) Relative expression of miR-221/222 between T47Dwt and T47DTamR exosomes. (b) Relative expression of miR-221/222 in T47Dwt cells treated with T47Dwt or T47DTamR exosomes respectively (TIFF 313 kb)

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Wei, Y., Lai, X., Yu, S. et al. Exosomal miR-221/222 enhances tamoxifen resistance in recipient ER-positive breast cancer cells. Breast Cancer Res Treat 147, 423–431 (2014). https://doi.org/10.1007/s10549-014-3037-0

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  • DOI: https://doi.org/10.1007/s10549-014-3037-0

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