Advertisement

Cell Biochemistry and Biophysics

, Volume 67, Issue 3, pp 1115–1125 | Cite as

DC Electric Fields Direct Breast Cancer Cell Migration, Induce EGFR Polarization, and Increase the Intracellular Level of Calcium Ions

  • Dan Wu
  • Xiuli Ma
  • Francis Lin
Original Paper

Abstract

Migration of cancer cells leads to invasion of primary tumors to distant organs (i.e., metastasis). Growing number of studies have demonstrated the migration of various cancer cell types directed by applied direct current electric fields (dcEF), i.e., electrotaxis, and suggested its potential implications in metastasis. MDA-MB-231 cell, a human metastatic breast cancer cell line, has been shown to migrate toward the anode of dcEF. Further characterizations of MDA-MB-231 cell electrotaxis and investigation of its underlying signaling mechanisms will lead to a better understanding of electrically guided cancer cell migration and metastasis. Therefore, we quantitatively characterized MDA-MB-231 cell electrotaxis and a few associated signaling events. Using a microfluidic device that can create well-controlled dcEF, we showed the anode-directing migration of MDA-MB-231 cells. In addition, surface staining of epidermal growth factor receptor (EGFR) and confocal microscopy showed the dcEF-induced anodal EGFR polarization in MDA-MB-231 cells. Furthermore, we showed an increase of intracellular calcium ions in MDA-MB-231 cells upon dcEF stimulation. Altogether, our study provided quantitative measurements of electrotactic migration of MDA-MB-231 cells, and demonstrated the electric field-mediated EGFR and calcium signaling events, suggesting their involvement in breast cancer cell electrotaxis.

Keywords

Breast cancer cell Electrotaxis EGFR Calcium Microfluidic device 

Abbreviations

dcEF

Direct current electric fields

ECM

Extracellular matrix

EGFR

Epidermal growth factor receptor

PDMS

Polydimethylsiloxane

EI

Electrotactic index

SEM

Standard error of the mean

MSD

Mean square displacement

Notes

Acknowledgments

This Research is supported by Grants from the Natural Sciences and Engineering Research Council of Canada (NSERC), the Canada Foundation for Innovation (CFI), the Manitoba Health Research Council (MHRC), and the University of Manitoba. We thank The Nano Systems Fabrication Laboratory (NSFL) at the University of Manitoba, and the Manitoba Centre for Proteomics and Systems Biology for research support. We thank Saravanan Nandagopal for helping collect chemical reagents, Jing Li and Jiandong Wu for helping with microfluidic device preparation. D.W. thanks MHRC for a postdoctoral fellowship.

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

12013_2013_9615_MOESM1_ESM.docx (298 kb)
Supplementary material 1 (DOCX 297 kb)

References

  1. 1.
    Matsubayashi, Y., Ebisuya, M., Honjoh, S., & Nishida, E. (2004). ERK activation propagates in epithelial cell sheets and regulates their migration during wound healing. Current Biology, 14, 731–735.PubMedCrossRefGoogle Scholar
  2. 2.
    McDougall, S., Dallon, J., Sherratt, J., & Maini, P. (2006). Fibroblast migration and collagen deposition during dermal wound healing: Mathematical modelling and clinical implications. Philosophical Transactions Series A, Mathematical, Physical, and Engineering Sciences, 364, 1385–1405.PubMedCrossRefGoogle Scholar
  3. 3.
    Ayala, R., Shu, T., & Tsai, L. H. (2007). Trekking across the brain: The journey of neuronal migration. Cell, 128, 29–43.PubMedCrossRefGoogle Scholar
  4. 4.
    Hatten, M. E. (2002). New directions in neuronal migration. Science, 297, 1660–1663.PubMedCrossRefGoogle Scholar
  5. 5.
    Keller, R. (2005). Cell migration during gastrulation. Current Opinion in Cell Biology, 17, 533–541.PubMedCrossRefGoogle Scholar
  6. 6.
    Friedl, P., & Wolf, K. (2003). Tumour-cell invasion and migration: Diversity and escape mechanisms. Nature Reviews Cancer, 3, 362–374.PubMedCrossRefGoogle Scholar
  7. 7.
    Yamaguchi, H., Wyckoff, J., & Condeelis, J. (2005). Cell migration in tumors. Current Opinion in Cell Biology, 17, 559–564.PubMedCrossRefGoogle Scholar
  8. 8.
    Kunkel, E. J., & Butcher, E. C. (2003). Plasma-cell homing. Nature Reviews Immunology, 3, 822–829.PubMedCrossRefGoogle Scholar
  9. 9.
    Lin, F., Nguyen, C., Wang, S., Saadi, W., Gross, S., & Jeon, N. (2005). Neutrophil migration in opposing chemoattractant gradients using microfluidic chemotaxis devices. Annals of Biomedical Engineering, 33, 475–482.PubMedCrossRefGoogle Scholar
  10. 10.
    Cooper, M., & Keller, R. (1984). Perpendicular orientation and directional migration of amphibian neural crest cells in DC electrical fields. Proceedings of the National Academy of Sciences USA, 81, 160–164.CrossRefGoogle Scholar
  11. 11.
    Tai, G., Reid, B., Cao, L., & Zhao, M. (2009). Electrotaxis and wound healing: Experimental methods to study electric fields as a directional signal for cell migration. Methods in Molecular Biology, 571, 77–97.PubMedCrossRefGoogle Scholar
  12. 12.
    McCaig, C., Rajnicek, A., Song, B., & Zhao, M. (2005). Controlling cell behavior electrically: Current views and future potential. Physiological Reviews, 85, 943–978.PubMedCrossRefGoogle Scholar
  13. 13.
    Mycielska, M., & Djamgoz, M. (2004). Cellular mechanisms of direct-current electric field effects: Galvanotaxis and metastatic disease. Journal of Cell Science, 117, 1631–1639.PubMedCrossRefGoogle Scholar
  14. 14.
    Robinson, K., & Messerli, M. (2003). Left/right, up/down: The role of endogenous electrical fields as directional signals in development, repair and invasion. Bioessays, 25, 759–766.PubMedCrossRefGoogle Scholar
  15. 15.
    Nuccitelli, R. (1988). Physiological electric fields can influence cell motility, growth and polarity. Advanced Cell Biology, 2, 21.Google Scholar
  16. 16.
    Djamgoz, M. B. A., Mycielska, M., Madeja, Z., Fraser, S., & Korohoda, W. (2001). Directional movement of rat prostate cancer cells in direct-current electric field: Involvement of voltagegated Na+ channel activity. Journal of Cell Science, 114, 2697–2705.PubMedGoogle Scholar
  17. 17.
    Yan, X., Han, J., Zhang, Z., Wang, J., Cheng, Q., Gao, K., et al. (2009). Lung cancer A549 cells migrate directionally in DC electric fields with polarized and activated EGFRs. Bioelectromagnetics, 30, 29–35.PubMedCrossRefGoogle Scholar
  18. 18.
    Pu, J., McCaig, C. D., Cao, L., Zhao, Z., Segall, J. E., & Zhao, M. (2007). EGF receptor signalling is essential for electric-field-directed migration of breast cancer cells. Journal of Cell Science, 120, 3395–3403.PubMedCrossRefGoogle Scholar
  19. 19.
    Price, J. E. (1996). Metastasis from human breast cancer cell lines. Breast Cancer Research and Treatment, 39, 93–102.PubMedCrossRefGoogle Scholar
  20. 20.
    Christofori, G. (2003). Changing neighbours, changing behaviour: Cell adhesion molecule-mediated signalling during tumour progression. EMBO Journal, 22, 2318–2323.PubMedCrossRefGoogle Scholar
  21. 21.
    Segall, J. E., Tyerech, S., Boselli, L., Masseling, S., Helft, J., Chan, A., et al. (1996). EGF stimulates lamellipod extension in metastatic mammary adenocarcinoma cells by an actin-dependent mechanism. Clinical & Experimental Metastasis, 14, 61–72.CrossRefGoogle Scholar
  22. 22.
    Levine, M. D., Liotta, L. A., & Stracke, M. L. (1995). Stimulation and regulation of tumor cell motility in invasion and metastasis. EXS, 74, 157–179.PubMedGoogle Scholar
  23. 23.
    Xue, C., Wyckoff, J., Liang, F., Sidani, M., Violini, S., Tsai, K. L., et al. (2006). Epidermal growth factor receptor overexpression results in increased tumor cell motility in vivo coordinately with enhanced intravasation and metastasis. Cancer Research, 66, 192–197.PubMedCrossRefGoogle Scholar
  24. 24.
    Hirsch, D. S., Shen, Y., & Wu, W. J. (2006). Growth and motility inhibition of breast cancer cells by epidermal growth factor receptor degradation is correlated with inactivation of Cdc42. Cancer Research, 66, 3523–3530.PubMedCrossRefGoogle Scholar
  25. 25.
    Zhao, M., Dick, A., Forrester, J., & McCaig, C. (1999). Electric field-directed cell motility involves up-regulated expression and asymmetric redistribution of the epidermal growth factor receptors and is enhanced by fibronectin and laminin. Molecular Biology of the Cell, 10, 1259–1276.PubMedCrossRefGoogle Scholar
  26. 26.
    Zhao, M., Pu, J., Forrester, J. V., & McCaig, C. D. (2002). Membrane lipids, EGF receptors, and intracellular signals colocalize and are polarized in epithelial cells moving directionally in a physiological electric field. FASEB J, 16, 857–859.PubMedGoogle Scholar
  27. 27.
    Fang, K. S., Ionides, E., Oster, G., Nuccitelli, R., & Isseroff, R. R. (1999). Epidermal growth factor receptor relocalization and kinase activity are necessary for directional migration of keratinocytes in DC electric fields. Journal of Cell Science, 112(Pt 12), 1967–1978.PubMedGoogle Scholar
  28. 28.
    Poo, M., & Robinson, K. (1977). Electrophoresis of concanavalin A receptors along embryonic muscle cell membrane. Nature, 265, 602–605.PubMedCrossRefGoogle Scholar
  29. 29.
    Brown, M. J., & Loew, L. M. (1994). Electric field-directed fibroblast locomotion involves cell surface molecular reorganization and is calcium independent. Journal of Cell Biology, 127, 117–128.PubMedCrossRefGoogle Scholar
  30. 30.
    Wu, D., & Lin, F. (2011). A receptor-electromigration-based model for cellular electrotactic sensing and migration. Biochemical and Biophysical Research Communications, 411, 695–701.PubMedCrossRefGoogle Scholar
  31. 31.
    Yang, S., & Huang, X. Y. (2005). Ca2+ influx through L-type Ca2+ channels controls the trailing tail contraction in growth factor-induced fibroblast cell migration. Journal of Biological Chemistry, 280, 27130–27137.PubMedCrossRefGoogle Scholar
  32. 32.
    Agle, K. A., Vongsa, R. A., & Dwinell, M. B. (2010). Calcium mobilization triggered by the chemokine CXCL12 regulates migration in wounded intestinal epithelial monolayers. Journal of Biological Chemistry, 285, 16066–16075.PubMedCrossRefGoogle Scholar
  33. 33.
    Prevarskaya, N., Skryma, R., & Shuba, Y. (2011). Calcium in tumour metastasis: New roles for known actors. Nature Reviews Cancer, 11, 609–618.PubMedCrossRefGoogle Scholar
  34. 34.
    Brundage, R. A., Fogarty, K. E., Tuft, R. A., & Fay, F. S. (1991). Calcium gradients underlying polarization and chemotaxis of eosinophils. Science, 254, 703–706.PubMedCrossRefGoogle Scholar
  35. 35.
    Hahn, K., DeBiasio, R., & Taylor, D. L. (1992). Patterns of elevated free calcium and calmodulin activation in living cells. Nature, 359, 736–738.PubMedCrossRefGoogle Scholar
  36. 36.
    Davis, F. M., Kenny, P. A., Soo, E. T., van Denderen, B. J., Thompson, E. W., Cabot, P. J., et al. (2011). Remodeling of purinergic receptor-mediated Ca2+ signaling as a consequence of EGF-induced epithelial–mesenchymal transition in breast cancer cells. PLoS ONE, 6, e23464.PubMedCrossRefGoogle Scholar
  37. 37.
    Raimondi, C., Chikh, A., Maffucci, T., & Falasca, M. (2012). A novel regulatory mechanism links PLCγ1 to PDK1. Journal of Cell Science, 125(Pt 13), 3153–3163.PubMedCrossRefGoogle Scholar
  38. 38.
    Onuma, E., & Hui, S. (1985). A calcium requirement for electric field-induced cell shape changes and preferential orientation. Cell Calcium, 6, 281–292.PubMedCrossRefGoogle Scholar
  39. 39.
    Hammerick, K. E., Longaker, M. T., & Prinz, F. B. (2010). In vitro effects of direct current electric fields on adipose-derived stromal cells. Biochemical and Biophysical Research Communications, 397, 12–17.PubMedCrossRefGoogle Scholar
  40. 40.
    Shanley, L., Walczysko, P., Bain, M., MacEwan, D., & Zhao, M. (2006). Influx of extracellular Ca2+ is necessary for electrotaxis in Dictyostelium. Journal of Cell Science, 119, 4741–4748.PubMedCrossRefGoogle Scholar
  41. 41.
    Li, J., Nandagopal, S., Wu, D., Romanuik, S. F., Paul, K., Thomson, D. J., et al. (2011). Activated T lymphocytes migrate toward the cathode of DC electric fields in microfluidic devices. Lab on a Chip, 11, 1298–1304.PubMedCrossRefGoogle Scholar
  42. 42.
    Wang, S. J., Saadi, W., Lin, F., Minh-Canh Nguyen, C., & Li Jeon, N. (2004). Differential effects of EGF gradient profiles on MDA-MB-231 breast cancer cell chemotaxis. Experimental Cell Research, 300, 180–189.PubMedCrossRefGoogle Scholar
  43. 43.
    Lin, F., & Butcher, E. (2006). T cell chemotaxis in a simple microfluidic device. Lab on a Chip, 6, 1462–1469.PubMedCrossRefGoogle Scholar
  44. 44.
    Lin, F., Baldessari, F., Gyenge, C. C., Sato, T., Chambers, R. D., Santiago, J. G., et al. (2008). Lymphocyte electrotaxis in vitro and in vivo. The Journal of Immunology, 181, 2465–2471.PubMedGoogle Scholar
  45. 45.
    Li, J., & Lin, F. (2011). Microfluidic devices for studying chemotaxis and electrotaxis. Trends in Cell Biology, 21, 489–497.PubMedCrossRefGoogle Scholar
  46. 46.
    Morita, M., Higuchi, C., Moto, T., Kozuka, N., Susuki, J., Itofusa, R., et al. (2003). Dual regulation of calcium oscillation in astrocytes by growth factors and pro-inflammatory cytokines via the mitogen-activated protein kinase cascade. Journal of Neuroscience, 23, 10944–10952.PubMedGoogle Scholar
  47. 47.
    Morita, M., & Kudo, Y. (2010). Growth factors upregulate astrocyte [Ca2+]i oscillation by increasing SERCA2b expression. Glia, 58, 1988–1995.PubMedCrossRefGoogle Scholar
  48. 48.
    Mishra, S., & Hamburger, A. W. (1993). Role of intracellular Ca2+ in the epidermal growth factor induced inhibition of protein tyrosine phosphatase activity in a breast cancer cell line. Biochemical and Biophysical Research Communications, 191, 1066–1072.PubMedCrossRefGoogle Scholar
  49. 49.
    Faupel, M., Vanel, D., Barth, V., Davies, R., Fentiman, I. S., Holland, R., et al. (1997). Electropotential evaluation as a new technique for diagnosing breast lesions. European Journal of Radiology, 24, 33–38.PubMedCrossRefGoogle Scholar
  50. 50.
    Li, J., Zhu, L., Zhang, M., & Lin, F. (2012). Microfluidic device for studying cell migration in single or co-existing chemical gradients and electric fields. Biomicrofluidics, 6, 24121–2412113.PubMedCrossRefGoogle Scholar
  51. 51.
    Minc, N., & Chang, F. (2010). Electrical control of cell polarization in the fission yeast Schizosaccharomyces pombe. Current Biology, 20, 710–716.PubMedCrossRefGoogle Scholar
  52. 52.
    Rezai, P., Siddiqui, A., Selvaganapathy, P. R., & Gupta, B. P. (2010). Electrotaxis of Caenorhabditis elegans in a microfluidic environment. Lab on a Chip, 10, 220–226.PubMedCrossRefGoogle Scholar
  53. 53.
    Even-Ram, S., & Yamada, K. M. (2005). Cell migration in 3D matrix. Current Opinion in Cell Biology, 17, 524–532.PubMedCrossRefGoogle Scholar
  54. 54.
    Sun, Y. S., Peng, S. W., Lin, K. H., & Cheng, J. Y. (2012). Electrotaxis of lung cancer cells in ordered three-dimensional scaffolds. Biomicrofluidics, 6, 14102–1410214.PubMedCrossRefGoogle Scholar
  55. 55.
    Nabeshima, K., Inoue, T., Shimao, Y., Kataoka, H., & Koono, M. (1999). Cohort migration of carcinoma cells: Differentiated colorectal carcinoma cells move as coherent cell clusters or sheets. Histology and Histopathology, 14, 1183–1197.PubMedGoogle Scholar
  56. 56.
    Li, L., Hartley, R., Reiss, B., Sun, Y., Pu, J., Wu, D., et al. (2012). E-cadherin plays an essential role in collective directional migration of large epithelial sheets. Cellular and Molecular Life Sciences, 69, 2779–2789.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Department of Physics and AstronomyUniversity of ManitobaWinnipegCanada
  2. 2.Department of ImmunologyUniversity of ManitobaWinnipegCanada
  3. 3.Department of Biological SciencesUniversity of ManitobaWinnipegCanada
  4. 4.Department of Biosystems EngineeringUniversity of ManitobaWinnipegCanada

Personalised recommendations