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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References
Fidler, I.J. Critical determinants of cancer metastasis: rationale for therapy. Cancer Chemother. Pharmacol. 43, S3–S10 (1999).
Ridley, A.J. Rho GTPases and cell migration. J. Cell Sci. 114, 2713–2722 (2001).
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).
Amano, M. et al. Formation of actin stress fibers and focal adhesions enhanced by Rho-kinase. Science 275, 1308–1311 (1997).
Kimura, K. et al. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase) Science 273, 245–248 (1996).
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).
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).
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).
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).
Fritz, G., Just, I. & Kaina, B. Rho GTPases are over-expressed in human tumors. Int. J. Cancer 81, 682–687 (1999).
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).
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).
Coleman, M.L. et al. Membrane blebbing during apoptosis results from caspase-mediated activation of ROCK I. Nature Cell Biol. 3, 339–345 (2001).
Uehata, M. et al. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature 389, 990–994 (1997).
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).
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).
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).
Insall, R.H. & Weiner, O.D. PIP3, PIP2, and cell movement — similar messages, different meanings? Dev. Cell 1, 743–747 (2001).
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).
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).
Cukierman, E., Pankov, R. & Yamada, K.M. Cell interactions with three-dimensional matrices. Curr. Opin. Cell Biol. 14, 633–639 (2002).
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).
Thiery, J.P. Epithelial–mesenchymal transitions in tumour progression. Nature Rev. Cancer 2, 442–454 (2002).
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).
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).
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).
Itoh, K. et al. An essential part for Rho-associated kinase in the transcellular invasion of tumor cells. Nature Med. 5, 221–225 (1999).
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).
Hidalgo, M. & Eckhardt, S.G. Development of matrix metalloproteinase inhibitors in cancer therapy. J. Natl Cancer Inst. 93, 178–193 (2001).
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).
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).
Sahai, E. & Marshall, C.J. ROCK and Dia have opposing effects on adherens junctions downstream of Rho. Nature Cell Biol. 4, 408–415 (2002).
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).
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).
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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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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DOI: https://doi.org/10.1038/ncb1019
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