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Multi-step pericellular proteolysis controls the transition from individual to collective cancer cell invasion

Nature Cell Biology volume 9, pages 893–904 (2007)Cite this article

Abstract

Invasive cell migration through tissue barriers requires pericellular remodelling of extracellular matrix (ECM) executed by cell-surface proteases, particularly membrane-type-1 matrix metalloproteinase (MT1-MMP/MMP-14). Using time-resolved multimodal microscopy, we show how invasive HT-1080 fibrosarcoma and MDA-MB-231 breast cancer cells coordinate mechanotransduction and fibrillar collagen remodelling by segregating the anterior force-generating leading edge containing β1 integrin, MT1-MMP and F-actin from a posterior proteolytic zone executing fibre breakdown. During forward movement, sterically impeding fibres are selectively realigned into microtracks of single-cell calibre. Microtracks become expanded by multiple following cells by means of the large-scale degradation of lateral ECM interfaces, ultimately prompting transition towards collective invasion similar to that in vivo. Both ECM track widening and transition to multicellular invasion are dependent on MT1-MMP-mediated collagenolysis, shown by broad-spectrum protease inhibition and RNA interference. Thus, invasive migration and proteolytic ECM remodelling are interdependent processes that control tissue micropatterning and macropatterning and, consequently, individual and collective cell migration.

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Figure 1: Enhanced collagenolysis and migration of HT-1080 cells in 3D collagen lattices after overexpression of MT1-MMP.
Figure 2: Distinct zones for adhesion, MT1-MMP location and collagenolysis in single migrating cells.
Figure 3: Real-time detection of collagen fibre cleavage and displacement.
Figure 4: Cell deformation after inhibition of pericellular collagenolysis.
Figure 5: Proteolytic and non-proteolytic migration in high-density collagen spheroid model.
Figure 6: Large-scale removal of ECM layers during multicellular invasion.
Figure 7: Requirement for pericellular collagenolysis in invasive multicellular but not single-cell migration.
Figure 8: Abrogation of collagenolytic track formation and collective invasion after knockdown of MT1-MMP in HT-MT1 cells.

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Acknowledgements

We thank M. Ott, M. Kuhn and A. Staudigel for excellent technical assistance; L. King for supplying COL2¾Cshort antibody; British Biotech Inc., UK, for supplying BB-2516; R. Fridman for supplying recombinant TIMP-1 and TIMP-2; E. Deryugina and A. Y. Strongin for providing HT-1080 cell lines; D. Pei for supplying human MT1-MMP cDNA; K. Müller-Hermelink for providing access to the confocal facility; and G. Krohne for providing access to electron microscopy. This work was supported by the Deutsche Forschungsgemeinschaft (FR 1155/7-1) and Deutsche Krebshilfe (AZ 106950). C.O. was supported by a Canada Research Chair Grant in Metalloproteinase Proteomics and Systems Biology, and E.T. by a CIHR Cell Signals Training Grant. S.S., Y.W. and Y.L. were supported by a research grant from NIH (RO1 CA86984).

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Author notes

  1. Yi I. Wu & Eric Tam

    Present address: Current addresses: Department of Pharmacology, University of North Carolina, Chapel Hill, North Carolina 27599, USA (Y.I.W.); Department of Protein Engineering, Genentech Inc., 1 DNA Way, South San Francisco, California 94080, USA (E.T.),

Authors and Affiliations

  1. DFG Research Center for Experimental Biomedicine and Department of Dermatology, Rudolf Virchow Center, University of Würzburg, Josef-Schneider-Strasse 2, Würzburg, 97080, Germany

    Katarina Wolf & Peter Friedl

  2. Department of Cell & Molecular Biology, Northwestern University Feinberg Medical School, 303 E. Superior Street, Lurie 3-111, Chicago, 60611, Illinois, USA

    Yi I. Wu, Yueying Liu & M. Sharon Stack

  3. Institute of Clinical Biochemistry and Pathobiochemistry, University of Würzburg, Grombuehlstrasse 12, Würzburg, 97080, Germany

    Jörg Geiger

  4. Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, V6T 1Z3, British Columbia, Canada

    Eric Tam & Christopher Overall

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Contributions

P.F. and K.W. conceived and designed the experiments. K.W., Y.W., Y.L. and E.T. performed the experiments. K.W., P.F. and J.G. analysed the data. S.S., J.G. and C.O. contributed reagents, material and analysis tools. K.W. and P.F. wrote the paper. All authors read and corrected the manuscript.

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Correspondence to Peter Friedl.

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Wolf, K., Wu, Y., Liu, Y. et al. Multi-step pericellular proteolysis controls the transition from individual to collective cancer cell invasion. Nat Cell Biol 9, 893–904 (2007). https://doi.org/10.1038/ncb1616

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