Skip to main content

Cell Aggregation Assays

  • Protocol
  • First Online:
Book cover Metastasis Research Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1070))

Abstract

Invasion of carcinoma cells is the result of a disequilibrium between invasion promoter and invasion suppressor gene products (Mareel and Van Roy, Anticancer Res 6:419–435, 1986). The E-cadherin/catenin complex is the most potent invasion suppressor at the cell membrane of epithelioid cells (Duffy et al., J Pathol 214:283–293, 2008). This complex consists of E-cadherin, a transmembrane glycoprotein of 120 kDa, which is linked to the actin cytoskeleton via the catenins (Behrens et al., J Cell Biol 108:2435–2447, 1989). Downregulation of the complex is a common feature in invasive carcinoma cells, and has been recognized at several levels, ranging from genomic mutations to functional deficiencies of an apparently intact complex (Ozawa et al., Proc Natl Acad Sci USA 87:4246–4250, 1990). Cell aggregation assays have been set up to test the functionality of the complex in epithelioid tumor cells. Functional integrity of the complex is a prerequisite for cell–cell adhesion between epithelial cells, and measuring cell aggregation in vitro has thus become another elegant tool to study differences between invasive and noninvasive cell types.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Mareel MM, Van Roy FM (1986) Are oncogenes involved in invasion and metastasis? Anticancer Res 6:419–435

    PubMed  CAS  Google Scholar 

  2. Duffy MJ, McGowan PM, Gallagher WM (2008) Cancer invasion and metastasis: changing views. J Pathol 214:283–293

    Article  PubMed  CAS  Google Scholar 

  3. Behrens J, Mareel MM, Van Roy FM, Birchmeier W (1989) Dissecting tumour cell invasion: epithelial cells acquire invasive properties after the loss of uvomorulin-mediated cell–cell adhesion. J Cell Biol 108:2435–2447

    Article  PubMed  CAS  Google Scholar 

  4. Ozawa M, Ringwald M, Kemler R (1990) Uvomorulin–catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule. Proc Natl Acad Sci USA 87:4246–4250

    Article  PubMed  CAS  Google Scholar 

  5. Van Aken E, De Wever O, Correia da Rocha AS, Mareel M (2001) Defective E-cadherin/catenin complexes in human cancer. Virchows Arch 439:725–751

    PubMed  Google Scholar 

  6. Bracke ME, Vyncke BM, Bruyneel EA, Vermeulen SJ, De Bruyne GK, Van Larebeke NA, Vleminckx K, Van Roy FM, Mareel MM (1993) Insulin-like growth factor I activates the invasion suppressor function of E-cadherin in MCF-7 human mammary carcinoma cells in vitro. Br J Cancer 68:282–289

    Article  PubMed  CAS  Google Scholar 

  7. Noe V, Willems J, Vandekerckhove J, Roy FV, Bruyneel E, Mareel M (1999) Inhibition of adhesion and induction of epithelial cell invasion by HAV-containing E-cadherin-specific peptides. J Cell Sci 112:127–135

    PubMed  CAS  Google Scholar 

  8. Kadmon G, Kowitz A, Altevogt P, Schachner M (1990) The neural cell adhesion molecule N-CAM enhances L1-dependent cell–cell interactions. J Cell Biol 110:193–208

    Article  PubMed  CAS  Google Scholar 

  9. Beuthan J, Minet O, Helfmann J, Herrig M, Muller G (1996) The spatial variation of the refractive index in biological cells. Phys Med Biol 41:369–382

    Article  PubMed  CAS  Google Scholar 

  10. Young IT (1977) Proof without prejudice: use of the Kolmogorov-Smirnov test for the analysis of histograms from flow systems and other sources. J Histochem Cytochem 25:935–941

    Article  PubMed  CAS  Google Scholar 

  11. Vermeulen SJ, Bruyneel EA, Van Roy FM, Mareel MM, Bracke ME (1995) Activation of the E-cadherin/catenin complex in human MCF-7 breast cancer cells by all-trans-retinoic acid. Br J Cancer 72:1447–1453

    Article  PubMed  CAS  Google Scholar 

  12. Vermeulen SJ, Nollet F, Teugels E, Vennekens KM, Malfait F, Philippe J, Speleman F, Bracke ME, Van Roy FM, Mareel MM (1999) The alphaE-catenin gene (CTNNA1) acts as an invasion-suppressor gene in human colon cancer cells. Oncogene 18:905–915

    Article  PubMed  CAS  Google Scholar 

  13. Bracke ME, Van Larebeke NA, Vyncke BM, Mareel MM (1991) Retinoic acid modulates both invasion and plasma membrane ruffling of MCF-7 human mammary carcinoma cells in vitro. Br J Cancer 63:867–872

    Article  PubMed  CAS  Google Scholar 

  14. Bracke ME, Charlier C, Bruyneel EA, Labit C, Mareel MM, Castronovo V (1994) Tamoxifen restores the E-cadherin function in human breast cancer MCF-7/6 cells and suppresses their invasive phenotype. Cancer Res 54:4607–4609

    PubMed  CAS  Google Scholar 

  15. Charlier C, Bruyneel E, Lechanteur C, Bracke M, Mareel M, Castronovo V (1996) Enhancement of tamoxifen-induced E-cadherin function by Ca2+ channel antagonists in human breast cancer MCF7/6 cells. Eur J Pharmacol 317:413–416

    Article  PubMed  CAS  Google Scholar 

  16. Bracke ME, Bruyneel EA, Vermeulen SJ, Vennekens K, Van Marck V, Mareel MM (1994) Citrus flavonoid effect on tumour invasion and metastasis. Food Technol 48:121–124

    CAS  Google Scholar 

  17. Van Marck V, Stove C, Van Den Bossche K, Stove V, Paredes J, Vander HY, Bracke M (2005) P-cadherin promotes cell–cell adhesion and counteracts invasion in human melanoma. Cancer Res 65:8774–8783

    Article  PubMed  Google Scholar 

  18. Koutsouki E, Beeching CA, Slater SC, Blaschuk OW, Sala-Newby GB, George SJ (2005) N-cadherin-dependent cell–cell contacts promote human saphenous vein smooth muscle cell survival. Arterioscler Thromb Vasc Biol 25:982–988

    Article  PubMed  CAS  Google Scholar 

  19. Canonici A, Steelant W, Rigot V, Khomitch-Baud A, Boutaghou-Cherid H, Bruyneel E, Van RF, Garrouste F, Pommier G, Andre F (2008) Insulin-like growth factor-I receptor, E-cadherin and alpha v integrin form a dynamic complex under the control of alpha-catenin. Int J Cancer 122:572–582

    Article  PubMed  CAS  Google Scholar 

  20. Debruyne D, Mareel M, Vanhoecke B, Bracke M (2009) Cell aggregation on agar as an indicator for cell-matrix adhesion: effects of opioids. In Vitro Cell Dev Biol Anim 45:473–482

    Article  PubMed  CAS  Google Scholar 

  21. Lahaye M, Rochas C (1997) Chemical structure and physico-chemical properties of agar. Hydrobiologia 221:137–148

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media, New York

About this protocol

Cite this protocol

Debruyne, D., Boterberg, T., Bracke, M.E. (2014). Cell Aggregation Assays. In: Dwek, M., Schumacher, U., Brooks, S. (eds) Metastasis Research Protocols. Methods in Molecular Biology, vol 1070. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4614-8244-4_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-4614-8244-4_6

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4614-8243-7

  • Online ISBN: 978-1-4614-8244-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics