The cytotoxic effect of memantine and its effect on cytoskeletal proteins expression in metastatic breast cancer cell line

Document Type : Original Article


1 Applied Physiology Research Center, Cardiovascular Research Institute, Department of Physiology, Isfahan University of Medical Sciences, Isfahan, Iran

2 Isfahan Cardiovascular Research Center, Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, Iran


Objective(s):Breast cancer is an important leading cause of death from cancer. Stathmin and tau proteins are regulators of cell motility, and their overexpression is associated with the progression and bad prognosis of breast cancer. Memantine, an N-methyl-D-aspartate (NMDA) receptor antagonist, is the potential inhibitor of tau protein in neurons. This study determines the effect of memantine on breast cancer cell migration and proliferation, tau and stathmin gene expression in cancer cells and its synergistic effect with paclitaxel.  
Materials and Methods: The cell proliferation was evaluated by MTT assay and for this purpose, MCF-7 breast cancer cells were treated with various concentration of memantine (2, 20 and 100 μg/ml). Tau and stathmin mRNA expression was evaluated through quantitative real time RT-PCR method. The migration of cancer cells treated with memantine for 24 hr was compared to non-treated cells using an in vitro transmembrane migration assay.
Results: Incubation of breast cancer cells with memantine resulted in a dose dependent reduction in cell survival (P=0.0001). Paclitaxel (100 nM) showed synergistic effect with memantine (P=0.0001). Memantine significantly decreased tau and stathmin mRNA expression (by RT-PCR), so that 100 µmol/l of memantine decreased tau and stathmin expression by 46% (P=0.0341) and 33% (P=0.043), respectively. Migration of cells was also decreased by memantine (P=0.0001).
Conclusion: The presented data shows that memantine reduced mRNA levels of tau and stathmin proteins and also reduced cellular migration.


1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin 2011; 61: 69–90.
2. National Cancer Institute SEER Stat Fact Sheets: Female breast cancer 2013.
3. Hermann PC, Huber SL, Herrler T, Aicher A, Ellwart JW, Guba M, et al. Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 2007; 1: 313-323.
4. Goodsell DS. The molecular perspective: micro-tubules and the taxanes. Oncologist 2000; 5: 345-346.
5. Quintá HR, Galigniana NM, Erlejman AG, Lagadari M, Piwien-Pilipuk G, Galigniana MD. Management of cytoskeleton architecture by molecular chaperones and immunophilins. Cell Signal 2011; 23: 1907–1920.
6. Jordan MA, Wilson L. Microtubules as a target for anticancer drugs. Nat Rev Cancer 2004; 4: 253-265.
7. Yang X, Kandil D, Cosar EF, Khan A. Fibroepithelial tumors of the breast: pathologic and immunohisto-chemical features and molecular mechanisms. Arch Pathol Lab Med 2014; 138: 25-36.
8. Mistry S, Atweh GF. Role of stathmin in the regulation of the mitotic spindle: potential applications in cancer therapy. Mt Sinai J Med 2002; 69: 299–304.
9. Marklund U, Larsson N, Gradin HM, Brattsand G, Gullberg M. Oncoprotein 18 is a phosphorylation-responsive regulator of microtubule dynamics. Embo J 1996; 15:5290–5298.
10. Mistry S, Luo XN, Atweh GF. Transcriptional regulation of phosphoprotein p18 during monocytic differentiation of U937 leukemic cells. Cell Mol Biol Res 1995; 41:103–110.
11. Luo X, Mookerjee B, Ferrari A, Mistry S, Atweh GF. Regulation of phosphoprotein p18 in leukemic cells. Cell cycle regulated phosphorylation by p34cdc2 kinase. J Biol Chem 1994; 269: 10312–10318.
12. Brattsand G. Correlation of oncoprotein 18/stathmin expression in human breast cancer with established prognostic factors. Br J Cancer 2000; 83: 311–318.
13. Goedert M, Spillantini MG, Jakes R, Rutherford D, Crowther RA. Multiple isoforms of human microtubule-associated protein tau: sequences and localization in neurofibrillary tangles of Alzheimer's disease. Neuron 1989; 3: 519–526. 
14. Johnsen JI, Aurelio ON, Kwaja Z, Jorgensen GE, Pellegata NS, Plattner R et al. p53-mediated negative regulation of stathmin/Op18 expression is associated with G(2)/M cell-cycle arrest. Int J Cancer 2000; 88: 685–691.
15. Li ZH, Xiong QY, Tu JH, Gong Y, Qiu W, Zhang HQ, et al. Tau proteins expressions in advanced breast cancer and its significance in taxane-containing neoadjuvant chemotherapy. Med Oncol 2013; 30: 591.
16. Zhou J, Qian S, Li H, He W, Tan X, Zhang Q, et al. Predictive value of microtubule-associated protein Tau in patients with recurrent and metastatic breast cancer treated with taxane-containing palliative chemotherapy. Tumour Biol 2015; 36: 3941-3947.
17. Sontag E, Nunbhakdi-Craig V, Lee G, Bloom GS, Mumby MC. Regulation of the phosphorylation state and microtubule-binding activity of Tau by protein phosphatase 2A. Neuron 1996; 17: 1201-1207.
18. Noble W, Planel E, Zehr C, Olm V, Meyerson J, SulemanF, et al. Inhibition of glycogen synthase kinase-3 by lithium correlates with reduced tauopathy and degeneration in vivo. Proc Natl Acad Sci U S A 2005; 102: 6990-6995
19. Olivares D, Deshpande VK, Shi Y, Lahiri DK, Greig NH, Rogers JT, et al. N-methyl D-aspartate (NMDA) receptor antagonists and memantine treatment for Alzheimer's disease, vascular dementia and Parkinson's disease. Curr Alzheimer Res 2012; 9: 746-758.
20. Wu TY, Chen CP. Dual action of memantine in Alzheimer disease: a hypothesis. Taiwan J Obstet Gynecol 2009; 48: 273-277.
21. Degerman Gunnarsson M, Kilander L, Basun H, Lannfelt L. Reduction of phosphorylated tau during memantine treatment of Alzheimer's disease. Dement Geriatr Cogn Disord 2007; 24: 247-252.
22. Mowla SJ, Emadi Bayegi M, Ziaee SA, Nikpoor P. Evaluating expression and potential diagnostic and prognostic values of Survivin in bladder tumors: a preliminary report. Urol J 2005; 2: 141-147.
23. Kim SH, Sehrawat A, Singh SV. Notch2 activation by benzyl isothiocyanate impedes its inhibitory effect on breast cancer cell migration. Breast Cancer Res Treat 2012; 134: 1067–1079
24. Baquero MT, Lostritto K, Gustavson MD, Bassi KA, Appia F, Camp RL, et al. Evaluation of prognostic and predictive value of microtubule associated protein tau in two independent cohorts. Breast Cancer Res 2011; 13: R85.
25. Matrone MA, Whipple RA, Thompson K, Cho EH, Vitolo MI, Balzer EM, et al. Metastatic breast tumors express increased tau, which promotes microtentacle formation and the reattachment of detached breast tumor cells. Oncogene 2010; 29: 3217-3227.
26. Matrone MA, Whipple RA, Balzer EM, Martin SS. Microtentacles tip the balance of cytoskeletal forces in circulating tumor cells. Cancer Res 2010; 70: 7737-7741.
27. Matrone M, Whipple R, Balzer E, Cho E, Yoon J, Martin S. Metastasis-associated microtentacles are induced in detached and circulating breast tumor cells by expression of the microtubule-binding protein, Tau. Cancer Res 2009; 69: 55.
28. Curmi PA, Noguès C, Lachkar S, Carelle N, Gonthier MP, Sobel A et al. Overexpression of stathmin in breast carcinomas points out to highly proliferative tumours. Br J Cancer 2000; 82: 142-150.
29. Belletti B, Nicoloso MS, Schiappacassi M, Berton S, Lovat F, Wolf K, et al. Stathmin activity influences sarcoma cell shape, motility, and metastatic potential. Mol Biol Cell 2008; 19: 2003-2013.
30. Trovik J, Wik E, Stefansson IM, Marcickiewicz J, Tingulstad S, Staff AC, et al. Stathmin overexpression identifies high-risk patients and lymph node metastasis in endometrial cancer. Clin Cancer Res 2011; 17: 3368-3377.
31. Miceli C, Tejada A, Castaneda A, Mistry SJ. Cell cycle inhibition therapy that targets stathmin in in vitro and in vivo models of breast cancer. Cancer Gene Ther 2013; 20: 298-307.
32. Alli E, Yang JM, Hait WN. Silencing of stathmin induces tumor-suppressor function in breast cancer cell lines harboring mutant p53. Oncogene 2007; 26: 1003-1012.
33. Meng XL, Su D, Wang L, Gao Y, Hu YJ, Yang HJ, et al. Low expression of stathmin in tumor predicts high response to neoadjuvant chemotherapy with docetaxel-containing regimens in locally advanced breast cancer. Genet Test Mol Biomarkers 2012; 16: 689-694.
34. Alli E, Bash-Babula J, Yang JM, Hait WN. Effect of stathmin on the sensitivity to antimicrotubule drugs in human breast cancer. Cancer Res 2002; 62: 6864–6869.