Increased vimentin mRNA expression in MCF-7 breast cancer cell line after repeated endoxifen-treatment

Paramita Paramita, Melva Louisa, Nafrialdi Nafrialdi



Background: Epithelial mesenchymal transition (EMT) plays a significant role in the development of cancer cell resistance to drugs. Vimentin, a type III intermediate filament protein, is a marker of EMT. Vimentin's over-expression in cancer correlates well with increased tumor growth, change in cell shape and poor prognosis. Endoxifen is an active metabolite of tamoxifen  and has become a new potent agent in the treatment of breast cancer. This is a study that aimed to investigate the effect of endoxifen exposure with or without estradiol on cell viability, cell morphology and EMT progression through the analysis of vimentin mRNA expression after 4-week treatment.

Methods: Endoxifen, 100 nM or 1,000 nM, with or without beta-estradiol were given repeatedly to MCF-7 cells. Cells treated with dimethyl sulfoxide (DMSO) 0.001% were used as control. After 2- and 4-week exposure, the cells were counted, analyzed for mRNA vimentin expression, and observed for morphological changes.

Results: Compared to control, there were significant decreases in vimentin mRNA expressions in endoxifen and endoxifen+β-estradiol treated cells after 2-weeks, which then significantly increased after 4-week compared with the 2-week exposure. We found no change in morphology of MCF-7 cells.

Conclusion: Repeated exposure of endoxifen might induce EMT progression through increased expression of vimentin in MCF-7 breast cancer cell line.


endoxifen; EMT; vimentin

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  1. Wu X, Hawse JR, Subramaniam M, Goetz MP, Ingle JN, Spelsberg TC. The tamoxifen metabolite, endoxifen, is a potent antiestrogen that targets estrogen receptor alpha for degradation in breast cancer cells. Cancer Res. 2009;69(5):1722–7.
  2. Hawse JR, Subramaniam M, Cicek M, Wu X, Gingery A, Grygo SB, et al. Endoxifen's molecular mechanism of action are concentration dependent and different than that of other anti-estrogens. PLoS One. 2013;8(1):e54613.
  3. Hawse JR, Subramaniam M, Wu X, Negron V, Muzaffer C, Lingle WL, et al. Abstract PD05-11: development, characterization, and effective in vitro treatment of an endoxifen resistant breast cancer cell line. Cancer Res. 2010;70(24):1.
  4. Housman G, Byler S, Heerboth S, Lapinska K, Longacre M, Snyder N, et al. Drug resistance in cancer: an overview. Cancers. 2014;6(3):1769–92.
  5. Satelli A, Li S. Vimentin in cancer and its potential molecular target in cancer therapy. Cell Mol Life Sci. 2012;68(18):3033–46.
  6. Mendez MG, Kojima S, Goldman RD. Vimentin induces changes in cell shape, motility, and adhesion during the epithelial to mesenchymal transition. FASEB J. 2010;24(6):1838–51.
  7. Micalizzi DS, Farabaugh SM, Ford HL. Epithelial-mesenchymal transition in cancer: parallels between normal development and tumor progression. J Mammary Gland Biol Neoplasia. 2010;15(2):117–34.
  8. Savagner P. The epithelial-mesenchymal transition (EMT) phenomenon. Ann Oncol. 2010;21(7):vii89–92.
  9. Kalluri R, Weinberg RA. The basic of epithelial-mesenchymal transition. J Clin Invest. 2009;119(6):1420–8.
  10. Wang Y, Zhou BP. Epithelial-mesenchymal transition--a hallmark of breast cancer metastasis. Cancer Hallm. 2013;1(1):38–49.
  11. Voulgari A, Pintzas A. Epithelial-mesenchymal transition in cancer metastasis: mechanisms, markers, and strategies to overcome drug resistance in the clinic. Biochim Biophys Acta. 2009;1796(2):75–90.
  12. Kusinska RU, Kordek R, Pluciennik E, Bednarek AK, Piekarski JH, Potemski P. Does vimentin help to delineate the so-called 'basal type breast cancer'? J Exp Clin Cancer Res. 2009;28(1):118.
  13. N, Triaspolitica. "Kanker Payudara: Informasi, Penyebab, Gejala, Stadium Dan Pengobatan." Mau Nanya Dong Dok. N.p, 28 June 2017. Web. 30 June 2017. Retreived from :
  14. Yang J, Bielenberg DR, Rodig SJ, Doiron R, Clifton MC, Kung AL, et al. Lipocalin 2 promotes breast cancer progression. Proc Natl Acad Sci USA. 2009;106(10):3913–8.
  15. Folkerd EJ, Lønning PE, Dowsett M. Interpreting plasma estrogen levels in breast cancer: caution needed. J Clin Oncol. 2014;32(14):1396–400.
  16. Jamalzadeh L, Ghafoori H, Sariri R, Rabuti H, Nasirzade J, Hasani H, et al. Cytotoxic effects of some common organic solvents on MCF-7, RAW-264.7 and human umbilical vein endothelial cells. Avicenna J Med Biochem. 2016;4(1):e33453.
  17. Yan XD, Li M, Yuan Y, Mao N, Pan LY. Biological comparison of ovarian cancer resistant cell lines to cisplatin and Taxol by two different administrations. Oncol Rep. 2007;17(5):1163–9.
  18. Louie MC, McClellan A, Siewit C, Kawabata L. Estrogen receptor regulates E2F1 expression to mediate tamoxifen resistance. Mol Cancer Res. 2010;8(3):343–52.
  19. Carlet J, Jarlier V, Harbarth S, Voss A, Goosens H, Pittet D, et al. Ready for a world without antibiotics? The pensières antibiotic resistance call to action. Antimicrob Resist Infect Control. 2012;1(1):11.
  20. Dong W, Zhang H, Li J, Guan H, He L, Wang Z, et al. Estrogen induces metastatic potential of papillary thyroid cancer cells through estrogen receptor α and β. Int J Endocrinol. 2013;2013(941568):1–6.
  21. Jiménez-Salazar JE, Posadas-Rodríguez P, Lazzarini-Lechuga RC, Luna-López A, Zentella-Dehesa A, Gómez-Quiroz LE, et al. Membrane-initiated estradiol signaling of epithelial-mesenchymal transition-associated mechanisms through regulation of tight junctions in human breast cancer cells. Horm Cancer. 2014;5(3):161–73.
  22. Zhao G, Nie Y, Lv M, He L, Wang T, Hou Y. ERβ-mediated estradiol enhances epithelial mesenchymal transition of lung adenocarcinoma through increasing transcription of midkine. Mol Endocrinol. 2012;26(8):1304–15.

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