Cellular Oncology

, Volume 40, Issue 1, pp 63–76 | Cite as

Collective cell migration of thyroid carcinoma cells: a beneficial ability to override unfavourable substrates

  • Liudmila Lobastova
  • Dominik Kraus
  • Alexander Glassmann
  • Dilaware Khan
  • Christian Steinhäuser
  • Christina Wolff
  • Nadine Veit
  • Jochen Winter
  • Rainer ProbstmeierEmail author
Original Paper



Tumor cell invasion and metastasis are life threatening events. Invasive tumor cells tend to migrate as collective sheets. In the present in vitro study we aimed to (i) assess whether collective tumor cells gain benefits in their migratory potential compared to single cells and (ii) to identify its putative underlying molecular mechanisms.


The migratory potential of single and collective carcinoma cells was assessed using video time lapse microscopy and cell migration assays in the absence and presence of seven potential gap junction inhibitors or the Rac1 inhibitor Z62954982. The perturbation of gap junctions was assessed using a dye diffusion assay. In addition, LDH-based cytotoxicity and RT-PCR-based expression analyses were performed.


Whereas single breast, cervix and thyroid carcinoma cells were virtually immobile on unfavourable plastic surfaces, we found that they gained pronounced migratory capacities as collectives under comparable conditions. Thyroid carcinoma cells, that were studied in more detail, were found to express specific subsets of connexins and to form active gap junctions as revealed by dye diffusion analysis. Although all potential gap junction blockers suppressed intercellular dye diffusion in at least one of the cell lines tested, only two of them were found to inhibit collective cell migration and none of them to inhibit single cell migration. In the presence of the Rac1 inhibitor Z62954982 collective migration, but not single cell migration, was found to be reduced up to 20 %.


Our data indicate that collective migration enables tumor cells to cross otherwise unfavourable substrate areas. This capacity seems to be independent of intercellular communication via gap junctions, whereas Rac1-dependent intracellular signalling seems to be essential.


Collective cell migration Gap junction Rac Single cell migration Thyroid carcinoma 



This research was supported by the Deutsche Forschungsgemeinschaft (STE 552/4, STE 552/5; to C.S.) and EU EraNet NEURON (Project BrIE; to C.S.).

Compliance with ethical standards

Conflict of interest

None declared.

Supplementary material (511 kb)
Movie 1 Single cell migration of B-CPAP cells on untreated cell culture plastic. (MOV 511 kb) (661 kb)
Movie 2 Single cell migration of FTC-133 cells on a fibronectin-covered surface. (MOV 661 kb) (584 kb)
Movie 3 Single cell migration of FTC-133 cells on a laminin-covered surface. (MOV 583 kb) (2.6 mb)
Movie 4 Collective cell migration: Typical migration pattern of a FTC-133 cell sheet. (MOV 2619 kb) (1.9 mb)
Movie 5 Collective cell migration: Integration of an isolated FTC-133 cell into a migrating cell sheet. (MOV 1934 kb) (2 mb)
Movie 6 Collective cell migration: Emigration of a single B-CPAP cell out of a migrating cell sheet. (MOV 2069 kb) (703 kb)
Movie 7 Collective cell migration: Mobility potential of B-CPAP cell aggregates. (MOV 703 kb)


  1. 1.
    R. Paduch, The role of lymphangiogenesis and angiogenesis in tumor metastasis. Cell Oncol 39, 397–410 (2016)CrossRefGoogle Scholar
  2. 2.
    J. Odenthal, R. Takes, P. Friedl, Plasticity of tumor cell invasion: governance by growth factors and cytokines. Carcinogenesis (2016). doi: 10.1093/carcin/bgw098 PubMedGoogle Scholar
  3. 3.
    M. Canel, A. Serrels, M. C. Frame, V. G. Brunton, E-cadherin-integrin crosstalk in cancer invasion and metastasis. J Cell Sci 126, 393–401 (2013)CrossRefPubMedGoogle Scholar
  4. 4.
    V. Saisongkorh, A. Maiuthed, P. Chanvorachote, Nitric oxide increases the migratory activity of non-small cell lung cancer cells via AKT-mediated integrin αv and β1 upregulation. Cell Oncol 39, 449–462 (2016)CrossRefGoogle Scholar
  5. 5.
    P. Friedl, K. Wolf, Tumour-cell invasion and migration: diversity and escape mechanisms. Nat Rev Cancer 3, 362–374 (2003)CrossRefPubMedGoogle Scholar
  6. 6.
    M. Yilmaz, G. Christofori, Mechanisms of motility in metastasizing cells. Mol Cancer Res 8, 629–642 (2010)CrossRefPubMedGoogle Scholar
  7. 7.
    K. J. Cheung, A. J. Ewald, A collective route to metastasis: seeding by tumor cell clusters. Science 352, 167–169 (2016)CrossRefPubMedGoogle Scholar
  8. 8.
    A. Haeger, K. Wolf, M. M. Zegers, P. Friedl, Collective cell migration: guidance principles and hierarchies. Trends Cell Biol 25, 556–566 (2015)CrossRefPubMedGoogle Scholar
  9. 9.
    M. Kotini, R. Mayor, Connexins in migration during development and cancer. Dev Biol 401, 143–151 (2015)CrossRefPubMedGoogle Scholar
  10. 10.
    V. Su, A. F. Lau, Connexins: mechanisms regulating protein levels and intercellular communication. FEBS Lett 588, 1212–1220 (2014)CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    G. Söhl, K. Willecke, Gap junctions and the connexin protein family. Cardiovasc Res 62, 228–232 (2004)CrossRefPubMedGoogle Scholar
  12. 12.
    P. Bedner, C. Steinhäuser, M. Theis, Functional redundancy and compensation among members of gap junction protein families? Biochim Biophys Acta 181, 1971–1984 (2012)CrossRefGoogle Scholar
  13. 13.
    A. A. Khalil, P. Friedl, Determinants of leader cells in collective cell migration. Integr Biol (Camb) 2, 568–574 (2010)CrossRefGoogle Scholar
  14. 14.
    N. Yamaguchi, T. Mizutani, K. Kawabata, H. Haga, Leader cells regulate collective cell migration via Rac activation in the downstream signaling of integrin β1 and PI3K. Sci Rep 5, 7656 (2015)CrossRefPubMedGoogle Scholar
  15. 15.
    M. Parri, P. Chiarugi, Rac and rho GTPases in cancer cell motility control. Cell Commun Signal 8, 23 (2008). doi: 10.1186/1478-811X-8-23 CrossRefGoogle Scholar
  16. 16.
    A. J. Ridley, Rho GTPase signalling in cell migration. Curr Opin Cell Biol 36, 103–112 (2015)CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    M. M. Zegers, P. Friedl, Rho GTPases in collective cell migration. Small GTPases 5, e28997 (2014). doi: 10.4161/sgtp.28997 CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    L. Delys, V. Detours, B. Franc, G. Thomas, T. Bogdanova, M. Tronko, F. Libert, J. E. Dumont, C. Maenhaut, Gene expression and the biological phenotype of papillary thyroid carcinoma. Oncogene 26, 7894–7893 (2007)CrossRefPubMedGoogle Scholar
  19. 19.
    A. Glassmann, J. Winter, D. Kraus, N. Veit, R. Probstmeier, Pharmacological suppression of the Ras/MAPK pathway in thyroid carcinoma cells can provoke opposite effects on cell migration and proliferation: the appearance of yin-yang effects and the need of combinatorial treatments. Int J Oncol 45, 2587–2595 (2014)PubMedGoogle Scholar
  20. 20.
    D. Hecker, J. Kappler, A. Glassmann, K. Schilling, W. Alt, Image analysis of time-lapse movies - a precision control guided approach to correct motion artefacts. J Neurosci Methods 172, 67–73 (2008)CrossRefPubMedGoogle Scholar
  21. 21.
    A. Glassmann, K. Reichmann, B. Scheffler, M. Glas, N. Veit, R. Probstmeier, Pharmacological targeting of the constitutively activated MEK/MAPK-dependent signaling pathway in glioma cells inhibits cell proliferation and migration. Int J Oncol 39, 1567–1575 (2011)PubMedGoogle Scholar
  22. 22.
    M. Koval, S. A. Molina, J. M. Burt, Mix and match: investigating heteromeric and heterotypic gap junction channels in model systems and native tissues. FEBS Lett 588, 1193–1204 (2014)CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    G. R. Juszczak, A. H. Swiergiel, Properties of gap junction blockers and their behavioural, cognitive and electrophysiological effects: animal and human studies. Prog Neuro-Psychopharmacol Biol Psychiatry 3, 181–198 (2009)CrossRefGoogle Scholar
  24. 24.
    G. Orend, R. Chiquet-Ehrismann, Adhesion modulation by antiadhesive molecules of the extracellular matrix. Exp Cell Res 261, 104–110 (2000)CrossRefPubMedGoogle Scholar
  25. 25.
    T. N. Wight, Versican: a versatile extracellular matrix proteoglycan in cell biology. Curr Opin Cell Biol 14, 617–623 (2002)CrossRefPubMedGoogle Scholar
  26. 26.
    B. W. Kiernan, B. Götz, A. Faissner, C. ffrench-Constant, Tenascin-C inhibits oligodendrocyte precursor cell migration by both adhesion-dependent and adhesion-independent mechanisms. Mol Cell Neurosci 7, 322–335 (1996)CrossRefPubMedGoogle Scholar
  27. 27.
    M. P. Burnham, P. M. Sharpe, C. Garner, R. Hughes, C. E. Pollard, J. Bowes, Investigation of connexin 43 uncoupling and prolongation of the cardiac QRS complex in preclinical and marketed drugs. Br J Pharmacol 171, 4808–4819 (2014)CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    K. Jensen, A. Patel, J. Klubo-Gwiezdzinska, A. Bauer, V. Vasko, Inhibition of gap junction transfer sensitizes thyroid cancer cells to anoikis. Endocr Relat Cancer 18, 613–626 (2011)CrossRefPubMedGoogle Scholar
  29. 29.
    Q. Aftab, W. C. Sin, C. C. Naus, Reduction in gap junction intercellular communication promotes glioma migration. Oncotarget 6, 11447–11464 (2015)CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    R. Oliveira, C. Christov, J. S. Guillamo, S. de Boüard, S. Palfi, L. Venance, M. Tardy, M. Peschanski, Contribution of gap junctional communication between tumor cells and astroglia to the invasion of the brain parenchyma by human glioblastomas. BMC Cell Biol 6, 7 (2005)CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    L. Li, R. Hartley, B. Reiss, Y. Sun, J. Pu, D. Wu, F. Lin, T. Hoang, S. Yamada, J. Jiang, M. Zhao, E-cadherin plays an essential role in collective directional migration of large epithelial sheets. Cell Mol Life Sci 69, 2779–2789 (2012)CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    O. Ramos Gde, L. Bernardi, I. Lauxen, M. Sant’Ana Filho, A. R. Horwitz, M. L. Lamers, Fibronectin modulates cell adhesion and signaling to promote single cell migration of highly invasive oral squamous cell carcinoma. PLoS One 11, e0151338 (2016). doi: 10.1371/journal.pone.0151338 CrossRefPubMedGoogle Scholar

Copyright information

© International Society for Cellular Oncology 2016

Authors and Affiliations

  • Liudmila Lobastova
    • 1
  • Dominik Kraus
    • 2
  • Alexander Glassmann
    • 3
  • Dilaware Khan
    • 4
  • Christian Steinhäuser
    • 4
  • Christina Wolff
    • 1
  • Nadine Veit
    • 1
  • Jochen Winter
    • 5
  • Rainer Probstmeier
    • 1
    Email author
  1. 1.Neuro- and Tumor Cell Biology Group, Department of Nuclear MedicineUniversity Hospital BonnBonnGermany
  2. 2.Department of Prosthodontics, Preclinical Education, and Material ScienceUniversity of BonnBonnGermany
  3. 3.Life Science InkubatorBonnGermany
  4. 4.Institute of Cellular Neurosciences, Medical FacultyUniversity of BonnBonnGermany
  5. 5.Oral Cell Biology Group, Department of Periodontology, Operative and Preventive DentistryUniversity of BonnBonnGermany

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