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Differing modes of tumour cell invasion have distinct requirements for Rho/ROCK signalling and extracellular proteolysis

Abstract

Rho family GTPases regulate the cytoskeleton and cell migration and are frequently overexpressed in tumours. Here, we identify two modes of tumour-cell motility in 3D matrices that involve different usage of Rho signalling. Rho signalling through ROCK promotes a rounded bleb-associated mode of motility that does not require pericellular proteolysis. This form of motility requires ezrin, which is localized in the direction of cell movement. In contrast, elongated cell motility is associated with Rac-dependent F-actin-rich protrusions and does not require Rho, ROCK or ezrin function. Combined blockade of extracellular proteases and ROCK negates the ability of tumour cells to switch between modes of motility and synergises to prevent tumour cell invasion.

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Figure 1: Different morphologies of tumour cells in a 3D matrix in vitro and in vivo.
Figure 5: Inhibition of extracellular proteases and ROCK cooperate to inhibit tumour cell invasion.
Figure 2: Different modes of motility have differential requirements for RhoA and ROCK signalling.
Figure 3: Time-lapse analysis of rounded cell motility.
Figure 4: Analysis of rounded cell morphology.

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References

  1. Fidler, I.J. Critical determinants of cancer metastasis: rationale for therapy. Cancer Chemother. Pharmacol. 43, S3–S10 (1999).

    Article  CAS  Google Scholar 

  2. Ridley, A.J. Rho GTPases and cell migration. J. Cell Sci. 114, 2713–2722 (2001).

    CAS  PubMed  Google Scholar 

  3. Ridley, A.J., Paterson, H.F., Johnston, C.L., Diekmann, D. & Hall, A. The small GTP-binding protein Rac regulates growth factor-induced membrane ruffling. Cell 70, 401–410 (1992).

    Article  CAS  Google Scholar 

  4. Amano, M. et al. Formation of actin stress fibers and focal adhesions enhanced by Rho-kinase. Science 275, 1308–1311 (1997).

    Article  CAS  Google Scholar 

  5. Kimura, K. et al. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase) Science 273, 245–248 (1996).

    Article  CAS  Google Scholar 

  6. Bretscher, A., Edwards, K. & Fehon, R.G. ERM proteins and merlin: integrators at the cell cortex. Nature Rev. Mol. Cell Biol. 3, 586–599 (2002).

    Article  CAS  Google Scholar 

  7. Yonemura, S., Matsui, T., Tsukita, S. & Tsukita, S. Rho-dependent and -independent activation mechanisms of ezrin/radixin/moesin proteins: an essential role for polyphosphoinositides in vivo. J. Cell Sci. 115, 2569–2580 (2002).

    CAS  PubMed  Google Scholar 

  8. Suwa, H. et al. Overexpression of the rhoC gene correlates with progression of ductal adenocarcinoma of the pancreas. Br. J. Cancer 77, 147–152 (1998).

    Article  CAS  Google Scholar 

  9. van Golen, K.L. et al. A novel putative low-affinity insulin-like growth factor-binding protein, LIBC (lost in inflammatory breast cancer), and RhoC GTPase correlate with the inflammatory breast cancer phenotype. Clin. Cancer Res. 5, 2511–2519 (1999).

    CAS  PubMed  Google Scholar 

  10. Fritz, G., Just, I. & Kaina, B. Rho GTPases are over-expressed in human tumors. Int. J. Cancer 81, 682–687 (1999).

    Article  CAS  Google Scholar 

  11. Clark, E.A., Golub, T.R., Lander, E.S. & Hynes, R.O. Genomic analysis of metastasis reveals an essential role for RhoC. Nature 406, 532–535 (2000).

    Article  CAS  Google Scholar 

  12. Kleinman, H.K. et al. Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. Biochemistry 21, 6188–6193 (1982).

    Article  CAS  Google Scholar 

  13. Coleman, M.L. et al. Membrane blebbing during apoptosis results from caspase-mediated activation of ROCK I. Nature Cell Biol. 3, 339–345 (2001).

    Article  CAS  Google Scholar 

  14. Uehata, M. et al. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature 389, 990–994 (1997).

    Article  CAS  Google Scholar 

  15. Decaudin, D. et al. Peripheral benzodiazepine receptor ligands reverse apoptosis resistance of cancer cells in vitro and in vivo. Cancer Res. 62, 1388–1393 (2002).

    CAS  PubMed  Google Scholar 

  16. Vial, E., Sahai, E. & Marshall, C.J. MAPK/Fra-1 inactivates β1 integrin signalling to Rho-GTP in tumour cells permitting Rac-dependent cell motility. Cancer Cell (in the press).

  17. Watton, S.R. & Downward, J. Akt/PKB localisation and 3′ phosphoinositide generation at sites of epithelial cell–matrix and cell–cell interaction. Curr. Biol. 9, 433–436 (1999).

    Article  CAS  Google Scholar 

  18. Insall, R.H. & Weiner, O.D. PIP3, PIP2, and cell movement — similar messages, different meanings? Dev. Cell 1, 743–747 (2001).

    Article  CAS  Google Scholar 

  19. Duden, R., Griffiths, G., Frank, R., Argos, P. & Kreis, T.E. β-COP, a 110 kd protein associated with non-clathrin-coated vesicles and the Golgi complex, shows homology to β-adaptin. Cell 64, 649–665 (1991).

    Article  CAS  Google Scholar 

  20. Crepaldi, T., Gautreau, A., Comoglio, P.M., Louvard, D. & Arpin, M. Ezrin is an effector of hepatocyte growth factor-mediated migration and morphogenesis in epithelial cells. J. Cell Biol. 138, 423–434 (1997).

    Article  CAS  Google Scholar 

  21. Cukierman, E., Pankov, R. & Yamada, K.M. Cell interactions with three-dimensional matrices. Curr. Opin. Cell Biol. 14, 633–639 (2002).

    Article  CAS  Google Scholar 

  22. Wolf, K. et al. Compensation mechanism in tumor cell migration: mesenchymal–amoeboid transition after blocking of pericellular proteolysis. J. Cell Biol. 160, 267–277 (2003).

    Article  CAS  Google Scholar 

  23. Thiery, J.P. Epithelial–mesenchymal transitions in tumour progression. Nature Rev. Cancer 2, 442–454 (2002).

    Article  CAS  Google Scholar 

  24. Hauck, C.R., Hsia, D.A., Ilic, D. & Schlaepfer, D.D. v-Src SH3-enhanced interaction with focal adhesion kinase at β1 integrin-containing invadopodia promotes cell invasion. J. Biol. Chem. 277, 12487–12490 (2002).

    Article  CAS  Google Scholar 

  25. Wyckoff, J.B., Jones, J.G., Condeelis, J.S. & Segall, J.E. A critical step in metastasis: in vivo analysis of intravasation at the primary tumor. Cancer Res. 60, 2504–2511 (2000).

    CAS  PubMed  Google Scholar 

  26. Wang, W. et al. Single cell behavior in metastatic primary mammary tumors correlated with gene expression patterns revealed by molecular profiling. Cancer Res. 62, 6278–6288 (2002).

    CAS  PubMed  Google Scholar 

  27. Itoh, K. et al. An essential part for Rho-associated kinase in the transcellular invasion of tumor cells. Nature Med. 5, 221–225 (1999).

    Article  CAS  Google Scholar 

  28. Takamura, M. et al. Inhibition of intrahepatic metastasis of human hepatocellular carcinoma by Rho-associated protein kinase inhibitor Y-27632. Hepatology 33, 577–581 (2001).

    Article  CAS  Google Scholar 

  29. Hidalgo, M. & Eckhardt, S.G. Development of matrix metalloproteinase inhibitors in cancer therapy. J. Natl Cancer Inst. 93, 178–193 (2001).

    Article  CAS  Google Scholar 

  30. Somlyo, A.V. et al. Rho kinase and matrix metalloproteinase inhibitors cooperate to inhibit angiogenesis and growth of human prostate cancer xenotransplants. FASEB J. 17, 223–234 (2003).

    Article  CAS  Google Scholar 

  31. Hill, C.S., Wynne, J. & Treisman, R. The Rho family GTPases RhoA, Rac1, and CDC42Hs regulate transcriptional activation by SRF. Cell 81, 1159–1170 (1995).

    Article  CAS  Google Scholar 

  32. Sahai, E. & Marshall, C.J. ROCK and Dia have opposing effects on adherens junctions downstream of Rho. Nature Cell Biol. 4, 408–415 (2002).

    Article  CAS  Google Scholar 

  33. Ishizaki, T. et al. p160ROCK, a Rho-associated coiled-coil forming protein kinase, works downstream of Rho and induces focal adhesions. FEBS Lett. 404, 118–124 (1997).

    Article  CAS  Google Scholar 

  34. Sahai, E., Olson, M.F. & Marshall, C.J. Cross-talk between Ras and Rho signalling pathways in transformation favours proliferation and increased motility. EMBO J. 20, 755–766 (2001).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank E. Vial and R. Lamb for advice and discussions, D. Bird for technical assistance and D. Croft for comments on the manuscript. E.S. and C.J.M. are funded by Cancer Research UK.

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Correspondence to Christopher J. Marshall.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information, Fig. S1 (PDF 1582 kb)

41556_2003_BFncb1019_MOESM2_ESM.avi

Movie 1. Time-lapse movie of GFP-A375m2 cells moving in 3D Matrigel, the first part of the movie shows a 360 rotation around the cells at a fixed timepoint before animation through the timepoints (timepoints are 450s apart). (AVI 3631 kb)

41556_2003_BFncb1019_MOESM3_ESM.avi

Movie 2. Time-lapse movie of GFP-A375m2 cells moving in 3D Matrigel, the movie shows a single confocal section (timepoints are 30 minutes apart). (AVI 62 kb)

41556_2003_BFncb1019_MOESM4_ESM.avi

Movie 3. Time-lapse movie of A375m2 cells on 2D tissue culture plastic in 10%DCS (timepoints are 5 minutes apart). (AVI 3295 kb)

41556_2003_BFncb1019_MOESM5_ESM.avi

Movie 4. Time-lapse movie of A375m2 cells on 2D tissue culture plastic in 10%DCS treated with 10µm Y27632 five hours prior to filming (timepoints are 5 minutes apart). (AVI 3364 kb)

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Sahai, E., Marshall, C. Differing modes of tumour cell invasion have distinct requirements for Rho/ROCK signalling and extracellular proteolysis. Nat Cell Biol 5, 711–719 (2003). https://doi.org/10.1038/ncb1019

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