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Pre-clinical effects of metformin and aspirin on the cell lines of different breast cancer subtypes

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Summary

Background Breast cancer is highly prevalent among women worldwide. It is classified into three main subtypes: estrogen receptor positive (ER+), human epidermal growth factor receptor 2 positive (HER2+), and triple negative breast cancer (TNBC). This study has evaluated the effects of aspirin and metformin, isolated or in a combination, in breast cancer cells of the different subtypes. Methods The breast cancer cell lines MCF-7, MDA-MB-231, and SK-BR-3 were treated with aspirin and/or metformin (0.01 mM - 10 mM); functional in vitro assays were performed. The interactions with the estrogen receptors (ER) were evaluated in silico. Results Metformin (2.5, 5 and 10 mM) altered the morphology and reduced the viability and migration of the ER+ cell line MCF-7, whereas aspirin triggered this effect only at 10 mM. A synergistic effect for the combination of metformin and aspirin (2.5, 5 or 10 mM each) was observed in the TNBC cell subtype MDA-MB-231, according to the evaluation of its viability and colony formation. Partial inhibitory effects were observed for either of the drugs in the HER2+ cell subtype SK-BR-3. The effects of metformin and aspirin partly relied on cyclooxygenase-2 (COX-2) upregulation, without the production of lipoxins. In silico, metformin and aspirin bound to the ERα receptor with the same energy. Conclusion We have provided novel evidence on the mechanisms of action of aspirin and metformin in breast cancer cells, showing favorable outcomes for these drugs in the ER+ and TNBC subtypes.

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Fig. 1: Metformin or aspirin induced a cell viability reduction.
Fig. 2: Metformin combined with aspirin reduced cell viability.
Fig. 3: Metformin and/or aspirin reduced colony formation.
Fig. 4: Metformin and/or aspirin did not induce cell migration.
Fig. 5: Metformin and/or aspirin reduced the migration of ER+ breast cancer cells.
Fig. 6: Metformin and/or aspirin increased the COX-2 protein expression.
Fig. 7: Metformin or aspirin bound to the active site of Estrogen Receptor Alpha (ERα).

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Acknowledgments

We would like to thank Dr. Eduardo Filippi-Chiela for providing the MCF-7 cell line and Dr. Mônica Ryff Moreira Roca Vianna for sharing the laboratory facilities for the western blotting analyzes.

Funding

This study was supported by grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brasil. MEAA is a master’s degree student in Cellular and Molecular Biology supported by the CAPES/PROEX Program. M.M.C. and W.F.A received grants from CNPq (Grant Numbers: 303842–2014-8 and 308883–2014-4, respectively).

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Correspondence to Maria Martha Campos.

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Amaral, M.E.A., Nery, L.R., Leite, C.E. et al. Pre-clinical effects of metformin and aspirin on the cell lines of different breast cancer subtypes. Invest New Drugs 36, 782–796 (2018). https://doi.org/10.1007/s10637-018-0568-y

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