Synergistic effects of combined treatment with simvastatin and exemestane on MCF‑7 human breast cancer cells

Shen,Yuanyuan; Du,Yingying; Zhang,Ying; Pan,Yueyin

doi:10.3892/mmr.2015.3406

Synergistic effects of combined treatment with simvastatin and exemestane on MCF‑7 human breast cancer cells

Authors:
- Yuanyuan Shen
- Yingying Du
- Ying Zhang
- Yueyin Pan
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Affiliations: Department of Oncology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, P.R. China, Department of Geriatrics, The Third Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230061, P.R. China
Published online on: March 3, 2015 https://doi.org/10.3892/mmr.2015.3406
Pages: 456-462

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Abstract

Breast cancer is associated with high levels of incidence, morbidity and mortality; therefore, the identification of effective chemopreventive strategies is crucial. It is important for clinicians to be able to identify the populations at risk who would benefit from chemoprevention, and the interventions that are effective and safe. The aim of the present study was to investigate the combined effects of simvastatin and exemestane on MCF‑7 human breast cancer cells. The anti‑proliferative effects of simvastatin and exemestane, alone and in combination, on the growth of MCF‑7 human breast cancer cells were assessed by MTT assay. The synergism between the two drugs was determined in vitro using the combination index (CI) analysis. Cell cycle distribution and apoptosis were analyzed by flow cytometry, and alterations to the signaling pathway in MCF‑7 cells were examined by immunoblotting following treatment with various regimens. The results of the MTT assay indicated that the combined treatment of simvastatin and exemestane significantly decreased the viability of MCF‑7 estrogen receptor‑positive (ER+) human breast cancer cells, as compared with those that were treated with the individual drugs (CI<1). In addition, coadministration of exemestane and simvastatin was shown to result in marked inhibition of tumor cell proliferation, significant cell cycle arrest at G0/G1 phase and induction of apoptosis, as compared with that of the control and individual drug‑treated cells. Furthermore, the results of the present study indicated that these synergistic effects may be associated with the B‑cell lymphoma 2 (Bcl‑2)/Bcl‑2‑associated X protein apoptotic pathway and the mitogen‑activated protein kinase/mammalian target of rapamycin/p70S6 kinase growth pathway. The combination of exemestane and simvastatin generated synergistic effects on MCF‑7 ER+ breast cancer cells, indicating that the combination of these drugs may be a potential therapeutic strategy for the treatment of hormone‑dependent breast cancer. The combination of the two inhibitors markedly increased the efficacy, as compared with the single‑agent treatment, suggesting that combination treatment could become a highly effective approach for breast cancer. The results of the present study suggested that this combination of drugs has therapeutic potential, and requires further mechanistic and biomarker investigations in clinical trials.

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1	Ferlay J, Parkin DM and Steliarova-Foucher E: Estimates of cancer incidence and mortality in Europe in 2008. Eur J Cancer. 46:765–781. 2010. View Article : Google Scholar : PubMed/NCBI
2	Chen S: An “omics” approach to determine the mechanisms of acquired aromatase inhibitor resistance. OMICS. 15:347–352. 2011. View Article : Google Scholar : PubMed/NCBI
3	Jankowitz RC and Davidson NE: Adjuvant endocrine therapy for breast cancer: how long is long enough? Oncology (Williston Park). 27:1210–1216. 2013.
4	Campagnoli C, Pasanisi P, Castellano I, Abba C, Brucato T and Berrino F: Postmenopausal breast cancer, androgens, and aromatase inhibitors. Breast Cancer Res Treat. 139:1–11. 2013. View Article : Google Scholar : PubMed/NCBI
5	Riemsma R, Forbes CA, Kessels A, et al: Systematic review of aromatase inhibitors in the first-line treatment for hormone sensitive advanced or metastatic breast cancer. Breast Cancer Res Treat. 123:9–24. 2010. View Article : Google Scholar : PubMed/NCBI
6	Li Z, Gu X, Tong J, et al: A meta-analysis of internal mammary lymph node metastasis in breast cancer patients. Onkologie. 36:747–752. 2013.PubMed/NCBI
7	Morris PG, McArthur HL and Hudis CA: Therapeutic options for metastatic breast cancer. Expert Opin Pharmacother. 10:967–981. 2009. View Article : Google Scholar : PubMed/NCBI
8	Rizzoli R, Body JJ, De Censi A, Reginster JY, Piscitelli P and Brandi ML; European Society for Clinical and Economical aspects of Osteoporosis and Osteoarthritis (ESCEO): Guidance for the prevention of bone loss and fractures in postmenopausal women treated with aromatase inhibitors for breast cancer: an ESCEO position paper. Osteoporos Int. 23:2567–2576. 2012. View Article : Google Scholar : PubMed/NCBI
9	Brodie A and Sabnis G: Adaptive changes result in activation of alternate signaling pathways and acquisition of resistance to aromatase inhibitors. Clin Cancer Res. 17:4208–4213. 2011. View Article : Google Scholar : PubMed/NCBI
10	Wong WW, Dimitroulakos J, Minden MD and Penn LZ: HMG-CoA reductase inhibitors and the malignant cell: the statin family of drugs as triggers of tumor-specific apoptosis. Leukemia. 16:508–519. 2002. View Article : Google Scholar : PubMed/NCBI
11	Superko HR, Momary KM and Li Y: Statins personalized. Med Clin North Am. 96:123–139. 2012. View Article : Google Scholar : PubMed/NCBI
12	Goldstein JL and Brown MS: Regulation of the mevalonate pathway. Nature. 343:425–430. 1990. View Article : Google Scholar : PubMed/NCBI
13	Liao JK: Isoprenoids as mediators of the biological effects of statins. J Clin Invest. 110:285–288. 2002. View Article : Google Scholar : PubMed/NCBI
14	Wejde J, Blegen H and Larsson O: Requirement for mevalonate in the control of proliferation of human breast cancer cells. Anticancer Res. 12:317–324. 1992.PubMed/NCBI
15	Spampanato C, De Maria S, Sarnataro M, et al: Simvastatin inhibits cancer cell growth by inducing apoptosis correlated to activation of Bax and down-regulation of BCL-2 gene expression. Int J Oncol. 40:935–941. 2012.
16	Kochuparambil ST, Al-Husein B, Goc A, Soliman S and Somanath PR: Anticancer efficacy of simvastatin on prostate cancer cells and tumor xenografts is associated with inhibition of Akt and reduced prostate-specific antigen expression. J Pharmacol Exp Ther. 336:496–505. 2011. View Article : Google Scholar
17	Campbell MJ, Esserman LJ, Zhou Y, et al: Breast cancer growth prevention by statins. Cancer Res. 66:8707–8714. 2006. View Article : Google Scholar : PubMed/NCBI
18	Cho SJ, Kim JS, Kim JM, Lee JY, Jung HC and Song IS: Simvastatin induces apoptosis in human colon cancer cells and in tumor xenografts, and attenuates colitis-associated colon cancer in mice. Int J Cancer. 123:951–957. 2008. View Article : Google Scholar : PubMed/NCBI
19	Nielsen SF, Nordestgaard BG and Bojesen SE: Statin use and reduced cancer-related mortality. N Engl J Med. 367:1792–1802. 2012. View Article : Google Scholar : PubMed/NCBI
20	Guterres FA, Martinez GR, Rocha ME and Winnischofer SM: Simvastatin rises reactive oxygen species levels and induces senescence in human melanoma cells by activation of p53/p21 pathway. Exp Cell Res. 319:2977–2988. 2013. View Article : Google Scholar : PubMed/NCBI
21	Cheng WH, Ho WY, Chang CF, et al: Simvastatin induces a central hypotensive effect via Ras-mediated signalling to cause eNOS up-regulation. Br J Pharmacol. 170:847–858. 2013. View Article : Google Scholar : PubMed/NCBI
22	Ahern TP, Pedersen L, Tarp M, et al: Statin prescriptions and breast cancer recurrence risk: a Danish nationwide prospective cohort study. J Natl Cancer Inst. 103:1461–1468. 2011. View Article : Google Scholar : PubMed/NCBI
23	Hu LX, Du YY, Zhang Y and Pan YY: Synergistic effects of exemestane and aspirin on MCF-7 human breast cancer cells. Asian Pac J Cancer Prev. 13:5903–5908. 2012. View Article : Google Scholar
24	Budman DR, Calabro A, Wang LG, et al: Synergism of cytotoxic effects of vinorelbine and paclitaxel in vitro. Cancer Invest. 18:695–701. 2000. View Article : Google Scholar : PubMed/NCBI
25	Budman DR and Calabro A: In vitro search for synergy and antagonism: evaluation of docetaxel combinations in breast cancer cell lines. Breast Cancer Res Treat. 74:41–46. 2002. View Article : Google Scholar : PubMed/NCBI
26	Chou TC and Talalay P: Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul. 22:27–55. 1984. View Article : Google Scholar : PubMed/NCBI
27	Paridaens RJ, Dirix LY, Beex LV, et al: Phase III study comparing exemestane with tamoxifen as first-line hormonal treatment of metastatic breast cancer in postmenopausal women: the European Organisation for Research and Treatment of Cancer Breast Cancer Cooperative Group. J Clin Oncol. 26:4883–4890. 2008. View Article : Google Scholar : PubMed/NCBI
28	Sestak I, Cuzick J, Sapunar F, et al ATAC Trialists’ Group: Risk factors for joint symptoms in patients enrolled in the ATAC trial: a retrospective, exploratory analysis. Lancet Oncol. 9:866–872. 2008. View Article : Google Scholar : PubMed/NCBI
29	Chen SH, Chou FF and Ko JY: The use of simvastatin with aromasin in an ovariectomized rat model: effects on the skeletal system. Chang Gung Med J. 33:509–514. 2010.PubMed/NCBI
30	Stoehr M, Mozet C, Boehm A, Aigner A, Dietz A and Wichmann G: Simvastatin suppresses head and neck squamous cell carcinoma ex vivo and enhances the cytostatic effects of chemotherapeutics. Cancer Chemother Pharmacol. 73:827–837. 2014. View Article : Google Scholar : PubMed/NCBI
31	Danilo C and Frank PG: Cholesterol and breast cancer development. Curr Opin Pharmacol. 12:677–682. 2012. View Article : Google Scholar : PubMed/NCBI
32	Llaverias G, Danilo C, Mercier I, et al: Role of cholesterol in the development and progression of breast cancer. Am J Pathol. 178:402–412. 2011. View Article : Google Scholar : PubMed/NCBI
33	Nelson ER, Wardell SE, Jasper JS, et al: 27-Hydroxycholesterol links hypercholesterolemia and breast cancer pathophysiology. Science. 342:1094–1098. 2013. View Article : Google Scholar : PubMed/NCBI
34	Imamura K, Ogura T, Kishimoto A, Kaminishi M and Esumi H: Cell cycle regulation via p53 phosphory-lation by a 5′-AMP activated protein kinase activator, 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside, in a human hepatocellular carcinoma cell line. Biochem Biophys Res Commun. 287:562–567. 2001. View Article : Google Scholar : PubMed/NCBI
35	Jones RG, Plas DR, Kubek S, et al: AMP-activated protein kinase induces a p53-dependent metabolic checkpoint. Mol Cell. 18:283–293. 2005. View Article : Google Scholar : PubMed/NCBI
36	Hadad SM, Baker L, Quinlan PR, et al: Histological evaluation of AMPK signalling in primary breast cancer. BMC Cancer. 9:3072009. View Article : Google Scholar : PubMed/NCBI
37	Bühler S and Laufer SA: p38 MAPK inhibitors: a patent review (2012 – 2013). Expert Opin Ther Pat. 24:535–554. 2014. View Article : Google Scholar
38	Misirkic M, Janjetovic K, Vucicevic L, et al: Inhibition of AMPK-dependent autophagy enhances in vitro antiglioma effect of simvastatin. Pharmacol Res. 65:111–119. 2012. View Article : Google Scholar
39	Yang PM, Liu YL, Lin YC, Shun CT, Wu MS and Chen CC: Inhibition of autophagy enhances anticancer effects of atorvastatin in digestive malignancies. Cancer Res. 70:7699–7709. 2010. View Article : Google Scholar : PubMed/NCBI
40	Woodard J, Sassano A, Hay N and Platanias LC: Statin-dependent suppression of the Akt/mammalian target of rapamycin signaling cascade and programmed cell death 4 up-regulation in renal cell carcinoma. Clin Cancer Res. 14:4640–4649. 2008. View Article : Google Scholar : PubMed/NCBI
41	Jaafar H, Abdullah S, Murtey MD and Idris FM: Expression of Bax and Bcl-2 in tumour cells and blood vessels of breast cancer and their association with angiogenesis and hormonal receptors. Asian Pac J Cancer Prev. 13:3857–3862. 2012. View Article : Google Scholar : PubMed/NCBI