Pre-clinical effects of metformin and aspirin on the cell lines of different breast cancer subtypes
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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.
KeywordsBreast cance Drug repurposing Metformin Aspirin
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.
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).
Compliance with ethical standards
Conflict of interest
None of the authors have any conflict of interest to disclose regarding the publication of the present manuscript.
This article has not featured any studies with human participants or animals whilst it was being performed by any of the authors.
- 1.American Cancer Society (2017) Cancer facts & figures 2017. American Cancer Society Inc., AtlantaGoogle Scholar
- 2.American Cancer Society (2016) Breast cancer facts & figures 2015–2016. American Cancer Society Inc., AtlantaGoogle Scholar
- 3.Orecchioni S, Reggiani F, Talarico G, Mancuso P, Calleri A, Gregato G, Labanca V, Noonan DM, Dallaglio K, Albini A, Bertolini F (2015) The biguanides metformin and phenformin inhibit angiogenesis, local and metastatic growth of breast cancer by targeting both neoplastic and microenvironment cells. Int J Cancer 136(6):E534–E544CrossRefPubMedGoogle Scholar
- 4.Wang J, Li G, Wang Y, Tang S, Sun X, Feng X, Li Y, Bao G, Li P, Mao X, Wang M, Liu P (2014) Suppression of tumor angiogenesis by metformin treatment via a mechanism linked to targeting of HER2/HIF-1α/VEGF secretion axis. Oncotarget 6(42):1–14Google Scholar
- 5.Talarico G, Orecchioni S, Dallaglio K, Reggiani F, Mancuso P, Calleri A, Gregato G, Labanca V, Rossi T, Noonan DM, Albini A, Bertolini F (2016) Aspirin and atenolol enhance metformin activity against breast cancer by targeting both neoplastic and microenvironment cells. Sci Rep 6(1):18673CrossRefPubMedPubMedCentralGoogle Scholar
- 14.van Staalduinen J, Frouws M, Reimers M, Bastiaannet E, van Herk-Sukel MPP, Lemmens V, de Steur WO, Hartgrink HH, van de Velde CJH, Liefers G-J (2016) The effect of aspirin and nonsteroidal anti-inflammatory drug use after diagnosis on survival of oesophageal cancer patients. Br J Cancer 114(9):1053–1059CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Cheng R, Liu Y, Cui J, Yang M, Liu X, Li P (2017) Aspirin regulation of c-myc and cyclinD1 proteins to overcome tamoxifen resistance in estrogen receptor-positive breast cancer cells. 8(18):30252–30264Google Scholar
- 19.Yue W, Zheng X, Lin Y, Yang CS, Xu Q, Carpizo D, Huang H, DiPaola RS, Tan X-L (2015) Metformin combined with aspirin significantly inhibit pancreatic cancer cell growth in vitro and in vivo by suppressing anti-apoptotic proteins Mcl-1 and Bcl-2. Oncotarget 6(25):21208–21224CrossRefPubMedPubMedCentralGoogle Scholar
- 23.With P, Stereochemistry R (2017) The crystal structure of aspirin acetylated human cyclooxygenase-2: insight into the formation of products with reversed stereochemistry. 55(8):1226–1238Google Scholar
- 25.Gobbetti T, Ducheix S, Le Faouder P, Perez T, Riols F, Boue J, Bertrand-Michel J, Dubourdeau M, Guillou H, Perretti M, Vergnolle N, Cenac N (2015) Protective effects of n-6 fatty acids-enriched diet on intestinal ischaemia/reperfusion injury involve lipoxin a4and its receptor. Br J Pharmacol 172(3):910–923CrossRefPubMedGoogle Scholar
- 28.Trott O, Olson AJ (2009) AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem 31(2):455–461Google Scholar
- 36.Kalinsky K, Zheng T, Hibshoosh H, Du X, Mundi P, Yang J, Refice S, Feldman SM, Taback B, Connolly E, Crew KD, Maurer MA, Hershman DL (2017) Proteomic modulation in breast tumors after metformin exposure: results from a ‘window of opportunity’ trial. Clin Transl Oncol 19(2):180–188CrossRefPubMedGoogle Scholar
- 37.Dowling RJO, Niraula S, Chang MC, Done SJ, Ennis M, McCready DR, Leong WL, Escallon JM, Reedijk M, Goodwin PJ, Stambolic V (2015) Changes in insulin receptor signaling underlie neoadjuvant metformin administration in breast cancer: a prospective window of opportunity neoadjuvant study. Breast Cancer Res 17(1):540CrossRefGoogle Scholar
- 38.Kim S, Shore DL, Wilson LE, Sanniez EI, Kim JH, Taylor JA, Sandler DP (2015) Lifetime use of nonsteroidal anti- inflammatory drugs and breast cancer risk : results from a prospective study of women with a sister with breast cancer. BMC Cancer 1–10Google Scholar
- 39.Elwood PC, Morgan G, Pickering JE, Galante J, Weightman AL, Morris D, Kelson M, Dolwani S (2016) Aspirin in the treatment of cancer: reductions in metastatic spread and in mortality: a systematic review and meta-analyses of published studies. PLoS One 11(4):e0152402CrossRefPubMedPubMedCentralGoogle Scholar
- 43.Cai X, Hu X, Tan X, Cheng W, Wang Q, Chen X, Guan Y, Chen C, Jing X (2015) Metformin induced AMPK activation, G0/G1 phase cell cycle arrest and the inhibition of growth of esophageal squamous cell carcinomas in vitro and in vivo. PLoS One 10(7):1–14Google Scholar
- 50.Chew GL, Huo CW, Huang D, Hill P, Cawson J, Frazer H, Hopper JL, Haviv I, Henderson MA, Britt K, Thompson EW (2015) Increased COX-2 expression in epithelial and stromal cells of high mammographic density tissues and in a xenograft model of mammographic density. Breast Cancer Res Treat 153(1):89–99CrossRefPubMedGoogle Scholar
- 51.Tury S, Becette V, Assayag F, Vacher S, Benoist C, Kamal M, Marangoni E, Bièche I, Lerebours F, Callens C (2016) Combination of COX-2 expression and PIK3CA mutation as prognostic and predictive markers for celecoxib treatment in breast cancer. Oncotarget 7(51):85124–85141CrossRefPubMedPubMedCentralGoogle Scholar
- 56.Duan Y, Chen F, Zhang A, Zhu B, Sun J, Xie Q, Chen Z (2014) Aspirin inhibits lipopolysaccharide-induced COX-2 expression and PGE2 production in porcine alveolar macrophages by modulating protein kinase C and protein tyrosine phosphatase activity. BMB Rep 47(1):45–50CrossRefPubMedPubMedCentralGoogle Scholar
- 64.Vethakanraj HS, Sesurajan BP, Padmanaban VP, Jayaprakasam M, Murali S, Sekar AK (2017) Anticancer effect of acid ceramidase inhibitor ceranib-2 in human breast cancer cell lines MCF-7, MDA MB-231 by the activation of SAPK/JNK, p38 MAPK apoptotic pathways, inhibition of the Akt pathway, downregulation of ERα. Anticancer Drugs 29(1):50–60CrossRefGoogle Scholar
- 66.Kim T, Kim HI, An JY, Lee J, Lee NR, Heo J, Kim JE, Yu J, Lee YS, Inn KS, Kim NJ (2016) Identification of novel estrogen receptor (ER) agonists that have additional and complementary anti-cancer activities via ER-independent mechanism. Bioorg Med Chem Lett 26(7):1844–1848CrossRefPubMedGoogle Scholar