Skip to main content
SpringerLink
Log in

Quantitative Method to Track Proteolytic Invasion in 3D Collagen

  • Protocol
  • First Online:
Pancreatic Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1882))

Abstract

Since many tumors are associated with a pronounced collagen-rich stromal reaction, there is increasing interest in understanding mechanisms by which cancer cells invade through the collagen barrier. Here we describe a quantitative method to track cell invasion in 3D collagen I gels. We analyze invasion by quantifying proteolytic tracks generated by invading cancer cells through a 3D collagen microenvironment. We provide a detailed protocol for this quantitative assay, which can be used to characterize signaling pathways that regulate invasion in the 3D microenvironment.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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. Friedl P, Alexander S (2011) Cancer invasion and the microenvironment: plasticity and reciprocity. Cell 147(5):992–1009. https://doi.org/10.1016/j.cell.2011.11.016

    Article  CAS  PubMed  Google Scholar 

  2. Friedl P, Locker J, Sahai E, Segall JE (2012) Classifying collective cancer cell invasion. Nat Cell Biol 14(8):777–783

    Article  Google Scholar 

  3. McDonald OG, Maitra A, Hruban RH (2012) Human correlates of provocative questions in pancreatic pathology. Adv Anat Pathol 19(6):351–362. https://doi.org/10.1097/PAP.0b013e318273f998

    Article  PubMed  PubMed Central  Google Scholar 

  4. Christiansen JJ, Rajasekaran AK (2006) Reassessing Epithelial to Mesenchymal Transition as a Prerequisite for Carcinoma Invasion and Metastasis. Cancer Res 66(17):8319–8326. https://doi.org/10.1158/0008-5472.can-06-0410

    Article  CAS  PubMed  Google Scholar 

  5. Hidalgo-Carcedo C, Hooper S, Chaudhry SI, Williamson P, Harrington K, Leitinger B, Sahai E (2011) Collective cell migration requires suppression of actomyosin at cell-cell contacts mediated by DDR1 and the cell polarity regulators Par3 and Par6. Nat Cell Biol 13(1):49–58 http://www.nature.com/ncb/journal/v13/n1/abs/ncb2133.html-supplementary-information

    Article  CAS  Google Scholar 

  6. Shields MA, Dangi-Garimella S, Redig AJ, Munshi HG (2012) Biochemical role of the collagen-rich tumor microenvironment in pancreatic cancer progression. Biochem J 441(2):541–552

    Article  CAS  Google Scholar 

  7. Chu GC, Kimmelman AC, Hezel AF, DePinho RA (2007) Stromal biology of pancreatic cancer. J Cell Biochem 101(4):887–907

    Article  CAS  Google Scholar 

  8. Egeblad M, Rasch MG, Weaver VM (2010) Dynamic interplay between the collagen scaffold and tumor evolution. Curr Opin Cell Biol 22(5):697–706. https://doi.org/10.1016/j.ceb.2010.08.015

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Maitra A, Hruban RH (2008) Pancreatic cancer. Annu Rev Pathol 3:157–188. https://doi.org/10.1146/annurev.pathmechdis.3.121806.154305

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Shields MA, Dangi-Garimella S, Krantz SB, Bentrem DJ, Munshi HG (2011) Pancreatic cancer cells respond to type i collagen by inducing snail expression to promote membrane type 1 matrix metalloproteinase-dependent collagen invasion. J Biol Chem 286(12):10495–10504. https://doi.org/10.1074/jbc.M110.195628

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Ottaviano AJ, Sun L, Ananthanarayanan V, Munshi HG (2006) Extracellular matrix-mediated membrane-type 1 matrix metalloproteinase expression in pancreatic ductal cells is regulated by transforming growth factor-beta1. Cancer Res 66(14):7032–7040

    Article  CAS  Google Scholar 

  12. Rowe RG, Weiss SJ (2009) Navigating ECM barriers at the invasive front: the cancer cell-stroma interface. Annu Rev Cell Dev Biol 25:567–595

    Article  CAS  Google Scholar 

  13. Hotary K, Allen E, Punturieri A, Yana I, Weiss SJ (2000) Regulation of cell invasion and morphogenesis in a three-dimensional type I collagen matrix by membrane-type matrix metalloproteinases 1, 2, and 3. J Cell Biol 149(6):1309–1323

    Article  CAS  Google Scholar 

  14. Hotary KB, Allen ED, Brooks PC, Datta NS, Long MW, Weiss SJ (2003) Membrane type I matrix metalloproteinase usurps tumor growth control imposed by the three-dimensional extracellular matrix. Cell 114(1):33–45

    Article  CAS  Google Scholar 

  15. Friedl P, Zanker KS, Brocker EB (1998) Cell migration strategies in 3-D extracellular matrix: differences in morphology, cell matrix interactions, and integrin function. Microsc Res Tech 43(5):369–378. https://doi.org/10.1002/(SICI)1097-0029(19981201)43:5<369::AID-JEMT3>3.0.CO;2-6

    Article  CAS  PubMed  Google Scholar 

  16. Cukierman E, Pankov R, Stevens DR, Yamada KM (2001) Taking cell-matrix adhesions to the third dimension. Science 294(5547):1708–1712. https://doi.org/10.1126/science.1064829

    Article  CAS  PubMed  Google Scholar 

  17. Griffith LG, Swartz MA (2006) Capturing complex 3D tissue physiology in vitro. Nat Rev Mol Cell Biol 7(3):211–224. https://doi.org/10.1038/nrm1858

    Article  CAS  PubMed  Google Scholar 

  18. Chow CR, Ebine K, Knab LM, Bentrem DJ, Kumar K, Munshi HG (2016) Cancer cell invasion in three-dimensional collagen is regulated differentially by Galpha13 protein and discoidin domain receptor 1-Par3 protein signaling. J Biol Chem 291(4):1605–1618. https://doi.org/10.1074/jbc.M115.669606

    Article  CAS  PubMed  Google Scholar 

  19. Wang X, Chow CR, Ebine K, Lee J, Rosner MR, Pan T, Munshi HG (2016) Interaction of tRNA with MEK2 in pancreatic cancer cells. Sci Rep 6:28260. https://doi.org/10.1038/srep28260

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Dangi-Garimella S, Redig AJ, Shields MA, Siddiqui MA, Munshi HG (2010) Rho-ROCK-myosin signaling mediates membrane type 1 matrix metalloproteinase-induced cellular aggregation of keratinocytes. J Biol Chem 285(36):28363–28372. https://doi.org/10.1074/jbc.M110.146019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Dangi-Garimella S, Strouch MJ, Grippo PJ, Bentrem DJ, Munshi HG (2011) Collagen regulation of let-7 in pancreatic cancer involves TGF-beta1-mediated membrane type 1-matrix metalloproteinase expression. Oncogene 30(8):1002–1008. https://doi.org/10.1038/onc.2010.485

    Article  CAS  PubMed  Google Scholar 

  22. Dangi-Garimella S, Krantz SB, Barron MR, Shields MA, Heiferman MJ, Grippo PJ, Bentrem DJ, Munshi HG (2011) Three-dimensional collagen i promotes gemcitabine resistance in pancreatic cancer through MT1-MMP-mediated expression of HMGA2. Cancer Res 71(3):1019–1028. https://doi.org/10.1158/0008-5472.CAN-10-1855

    Article  CAS  PubMed  Google Scholar 

  23. Wolf K, Wu YI, Liu Y, Geiger J, Tam E, Overall C, Stack MS, Friedl P (2007) Multi-step pericellular proteolysis controls the transition from individual to collective cancer cell invasion. Nat Cell Biol 9(8):893–904. https://doi.org/10.1038/ncb1616

    Article  CAS  PubMed  Google Scholar 

  24. Wolf K, Te Lindert M, Krause M, Alexander S, Te Riet J, Willis AL, Hoffman RM, Figdor CG, Weiss SJ, Friedl P (2013) Physical limits of cell migration: control by ECM space and nuclear deformation and tuning by proteolysis and traction force. J Cell Biol 201(7):1069–1084. https://doi.org/10.1083/jcb.201210152

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by R01CA186885 grant (to H.G. Munshi) from the NCI and a Merit award I01BX002922 (to H.G. Munshi) from the Department of Veterans Affairs.

Author information

Authors and Affiliations

  1. Department of Medicine, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA

    Kazumi Ebine, Christina R. Chow & Hidayatullah G. Munshi

  2. Jesse Brown VA Medical Center, Chicago, IL, USA

    Kazumi Ebine & Hidayatullah G. Munshi

  3. The Robert H. Lurie Comprehensive Cancer Center, Chicago, IL, USA

    Hidayatullah G. Munshi

Authors
  1. Kazumi Ebine
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christina R. Chow
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hidayatullah G. Munshi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hidayatullah G. Munshi .

Editor information

Editors and Affiliations

  1. Department of Pathology and Cell Biology, Columbia University Medical Center, New York, NY, USA

    Gloria H. Su

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Ebine, K., Chow, C.R., Munshi, H.G. (2019). Quantitative Method to Track Proteolytic Invasion in 3D Collagen. In: Su, G. (eds) Pancreatic Cancer. Methods in Molecular Biology, vol 1882. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8879-2_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8879-2_15

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8878-5

  • Online ISBN: 978-1-4939-8879-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation