Skip to main content

Basement Membrane Invasion Assays: Native Basement Membrane and Chemoinvasion Assay

  • Protocol
  • First Online:
Adhesion Protein Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1046))

Abstract

To escape the primary tumor and infiltrate stromal compartments, invasive cancer cells must traverse the basement membrane (BM). To break this dense matrix, cells develop finger-like protrusions, called invadopodia, at their ventral surface. Invadopodia secrete proteases to degrade the BM, and then elongate which allows the cell to invade the subjacent tissue. Here, we describe two complementary invasion assays. The native BM invasion assay, based on BM isolated from rat or mouse mesentery, is a physiologically significant approach for studying the stages of BM crossing at the cellular level. The Matrigel-based chemoinvasion assay is a powerful technique for studying invadopodia’s molecular composition and organization at the subcellular level.

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 139.00
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. Yurchenco PD (2011) Basement membranes: cell scaffoldings and signaling platforms. Cold Spring Harb Perspect Biol 3(2):1–27

    Google Scholar 

  2. Sherwood DR (2006) Cell invasion through basement membranes: an anchor of understanding. Trends Cell Biol 16(5):250–256

    Article  PubMed  CAS  Google Scholar 

  3. Carman CV, Sage PT, Sciuto TE, de la Fuente MA, Geha RS, Ochs HD, Dvorak HF, Dvorak AM, Springer TA (2007) Transcellular diapedesis is initiated by invasive podosomes. Immunity 26(6):784–797

    Article  PubMed  CAS  Google Scholar 

  4. Schoumacher M, Louvard D, Vignjevic D (2011) Cytoskeleton networks in basement membrane transmigration. Eur J Cell Biol 90(2–3):93–99

    Article  PubMed  CAS  Google Scholar 

  5. Rowe RG, Weiss SJ (2008) Breaching the basement membrane: who, when and how? Trends Cell Biol 18(11):560–574

    Article  PubMed  CAS  Google Scholar 

  6. Ihara S, Hagedorn EJ, Morrissey MA, Chi Q, Motegi F, Kramer JM, Sherwood DR (2011) Basement membrane sliding and targeted adhesion remodels tissue boundaries during uterine-vulval attachment in Caenorhabditis elegans. Nat Cell Biol 13(6):641–651

    Article  PubMed  CAS  Google Scholar 

  7. Schoumacher M, Goldman RD, Louvard D, Vignjevic DM (2010) Actin, microtubules, and vimentin intermediate filaments cooperate for elongation of invadopodia. J Cell Biol 189(3):541–556

    Article  PubMed  CAS  Google Scholar 

  8. Hotary K, Li XY, Allen E, Stevens SL, Weiss SJ (2006) A cancer cell metalloprotease triad regulates the basement membrane transmigration program. Genes Dev 20(19):2673–2686

    Article  PubMed  CAS  Google Scholar 

  9. Artym VV, Yamada KM, Mueller SC (2009) ECM degradation assays for analyzing local cell invasion. Methods Mol Biol 522: 211–219

    Article  PubMed  CAS  Google Scholar 

  10. Poincloux R, Lizarraga F, Chavrier P (2009) Matrix invasion by tumour cells: a focus on MT1-MMP trafficking to invadopodia. J Cell Sci 122(Pt 17):3015–3024

    Article  PubMed  CAS  Google Scholar 

  11. Lizarraga F, Poincloux R, Romao M, Montagnac G, Le Dez G, Bonne I, Rigaill G, Raposo G, Chavrier P (2009) Diaphanous-related formins are required for invadopodia formation and invasion of breast tumor cells. Cancer Res 69(7):2792–2800

    Article  PubMed  CAS  Google Scholar 

  12. Geraldo S, Simon A, Elkhatib N, Louvard D, Fetler L, Vignjevic DM (2012) Do cancer cells have distinct adhesions in 3D collagen matrices and in vivo? Eur J Cell Biol 91(11–12): 930–937

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work is supported by Association pour la Recherche sur le Cancer (ARC-SFI12111203862) and ANR-09-JCJC-0023-01.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Schoumacher, M., Glentis, A., Gurchenkov, V.V., Vignjevic, D.M. (2013). Basement Membrane Invasion Assays: Native Basement Membrane and Chemoinvasion Assay. In: Coutts, A. (eds) Adhesion Protein Protocols. Methods in Molecular Biology, vol 1046. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-538-5_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-538-5_8

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-537-8

  • Online ISBN: 978-1-62703-538-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics