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The miR-21/PTEN/Akt signaling pathway is involved in the anti-tumoral effects of zoledronic acid in human breast cancer cell lines

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Abstract

Preclinical data indicate a direct anti-tumor effect of zoledronic acid (ZA) outside the skeleton, but its molecular mechanism is still not completely clarified. The aim of this study was to investigate the anti-cancer effects of ZA in human breast cancer cell lines, suggesting that they may in part be mediated via the miR-21/PTEN/Akt signaling pathway. The effect of ZA on cell viability was measured by MTT assay, and cell death induction was analyzed using either a double AO/EtBr staining and M30 ELISA assay. A Proteome Profiler Human Apoptosis Array was executed to evaluate the molecular basis of ZA-induced apoptosis. Cell cycle analysis was executed by flow cytometry. The effect of ZA on miR-21 expression was quantified by qRT-PCR, and the amount of PTEN protein and its targets were analyzed by Western blot. ZA inhibited cell growth in a concentration- and time-dependent manner, through the activation of cell death pathways and arrest of cell cycle progression. ZA downregulated the expression of miR-21, resulting in dephosphorilation of Akt and Bad and in a significant increase of p21 and p27 proteins expression. These results were observed also in MDA-MB-231 cells, commonly used as an experimental model of bone metastasis of breast cancer. This study revealed, for the first time, an involvement of the miR-21/PTEN/Akt signaling pathway in the mechanism of ZA anti-cancer actions in breast cancer cells. We would like to underline that this pathway is present both in the hormone responsive BC cell line (MCF-7) as well as in a triple negative cell line (MDA-MB-231). Taken together these results reinforce the use of ZA in clinical practice, suggesting the role of miR-21 as a possible mediator of its therapeutic efficacy.

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Acknowledgments

This work was supported by A.O. Istituti Ospitalieri di Cremona, Cremona (Italy) and by ARCO, Associazione Ricerca in Campo Oncologico, Cremona (Italy). Zoledronic Acid (Zometa®) was kindly given by Novartis Pharmaceuticals.

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Authors and Affiliations

  1. Section of Pharmacology, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11, 25123, Brescia, Italy

    M. Fragni, S. A. Bonini, P. Bettinsoli, S. Bodei, P. F. Spano, M. Memo & S. Sigala

  2. Department of Medical, Surgery and Health Sciences, University of Trieste, Piazza Ospitale 1, 34129, Trieste, Italy

    D. Generali

  3. Breast Unit and Molecular Therapy and Pharmacogenomic Unit, Istituti Ospitalieri di Cremona, Viale Concordia 1, Cremona, Italy

    A. Bottini

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  1. M. Fragni
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Correspondence to M. Fragni.

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Fragni, M., Bonini, S.A., Bettinsoli, P. et al. The miR-21/PTEN/Akt signaling pathway is involved in the anti-tumoral effects of zoledronic acid in human breast cancer cell lines. Naunyn-Schmiedeberg's Arch Pharmacol 389, 529–538 (2016). https://doi.org/10.1007/s00210-016-1224-8

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