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An essential part for Rho–associated kinase in the transcellular invasion of tumor cells

Abstract

Adhesion of tumor cells to host cell layers and subsequent transcellular migration are pivotal steps in cancer invasion and metastasis1,2,3. The small GTPase Rho controls cell adhesion and motility through reorganization of the actin cytoskeleton and regulation of actomyosin contractility4. Cultured rat MM1 hepatoma cells migrate through a mesothelial cell monolayer in vitro in a serum–dependent, Rho–mediated manner5. Among several proteins isolated as putative target molecules of Rho, the ROCK (ROK) family of Rho–associated serine–threonine protein kinases6,7,8 are thought to participate in the induction of focal adhesions and stress fibers in cultured cells9, and to mediate calcium sensitization of smooth muscle contraction by enhancing phosphorylation of the regulatory light chain of myosin10. Transfection of MM1 cells with cDNA encoding a dominant active mutant of ROCK conferred invasive activity independently of serum and Rho. In contrast, expression of a dominant negative, kinase–defective ROCK mutant substantially attenuated the invasive phenotype. A specific ROCK inhibitor (Y–27632; ref. 11) blocked both Rho–mediated activation of actomyosin and invasive activity of these cells. Furthermore, continuous delivery of this inhibitor using osmotic pumps considerably reduced the dissemination of MM1 cells implanted into the peritoneal cavity of syngeneic rats. These results indicate that ROCK plays an essential part in tumor cell invasion, and demonstrate its potential as a therapeutic target for the prevention of cancer invasion and metastasis.

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Figure 1: Effects of expression of p160ROCK mutants or active Rho on the morphology and invasive activity of MM1 cells in vitro.
Figure 2: a–b, In vitro invasion of transfected cells.
Figure 3: Effects of ROCK inhibitors on the in vitro invasiveness, MLC20 phosphorylation and adherence to the plastic dish of MM1 cells.
Figure 4: Effect of Y–27632 on the peritoneal dissemination of MM1 cells.

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References

  1. Fidler, I. J. Cancer metastasis. Br. Med. Bull. 47, 157–177 (1991).

    Article  CAS  Google Scholar 

  2. Nicolson, G.L. et al. Adhesive, invasive, and growth properties of selected metastatic variants of a murine large–cell lymphoma. Invasion Metastasis 9, 102–116 (1989).

    CAS  PubMed  Google Scholar 

  3. Liotta, L.A., Steeg, P.S. & Stetler–Stevenson, W.G. Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell 64, 327–336 (1991).

    Article  CAS  Google Scholar 

  4. Chrzanowska–Wodnicka, M. & Burridge, K. Rho–stimulated contractility drives the formation of stress fibers and focal adhesions. J. Cell Biol. 133, 1403–1415 (1996).

    Article  Google Scholar 

  5. Yoshioka, K., Matsumura, F., Akedo, H. & Itoh, K. Small GTP–binding protein Rho stimulates the actomyosin system, leading to invasion of tumor cells J. Biol. Chem. 273, 5146– 5154 (1998).

    Article  CAS  Google Scholar 

  6. Leung, T., Manser, E., Tan, L. & Lim, L. A novel serine/threonine kinase binding the Ras–related RhoA GTPase which translocates the kinase to peripheral membranes. J. Biol. Chem. 270, 29051–29054 (1995).

    Article  CAS  Google Scholar 

  7. Ishizaki, T. et al. The small GTP–binding protein Rho binds to and activates a 160 kDa Ser/Thr protein kinase homologous to myotonic dystrophy kinase. EMBO J. 15, 1885–1893 (1996).

    Article  CAS  Google Scholar 

  8. Matsui, T. et al. Rho–associated kinase, a novel serine/threonine kinase, as a putative target for the small GTP binding protein Rho. EMBO J. 15, 2208–2216 (1996).

    Article  CAS  Google Scholar 

  9. 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).

  10. Amano, M. et al. Phosphorylation and activation of myosin by Rho–associated kinase (Rho–kinase). J. Biol. Chem. 271, 20246–20249 (1996).

    Article  CAS  Google Scholar 

  11. Uehata, M. et al. A key role for p160ROCK–mediated Ca++ sensitization of smooth muscle in hypertension. Nature 389, 990–994 (1997).

    Article  CAS  Google Scholar 

  12. Sekine, A., Fujiwara, M. & Narumiya, S. Asparagine residue in the rho gene product is the modification site for Botulinum ADP–ribosyltransferase. J. Biol. Chem. 264, 8602–8605 (1989).

    CAS  PubMed  Google Scholar 

  13. Sahai, E., Alberts, A. S. & Treisman, R. RhoA effector mutants reveal distinct effector pathway for cytoskeletal reorganization, SRF activation and transformation. EMBO J. 17, 1350–1361 (1998).

    Article  CAS  Google Scholar 

  14. Matsumura, F., Ono, S., Yamakita, Y., Totsukawa, G. & Yamashiro, S. Specific localization of serine 19 phosphorylated myosin II during cell locomotion and mitosis of cultured cells. J. Cell Biol. 140, 119–129 (1998).

    Article  CAS  Google Scholar 

  15. Vleminckx, K., Vakaet, L., Mareel, M., Fiers, W. & Van Roy, F. Genetic manipulation of E–cadherin expression by epithelial tumor cells reveals an invasion suppressor role. Cell 66, 107–119 (1991).

    Article  CAS  Google Scholar 

  16. Perl, A., Wilgenbus, P., Dahl, U., Semb, H. & Christofori, G. A causal role for E–cadherin in the transition from adenoma to carcinoma. Nature 392, 190–193 (1998).

    Article  CAS  Google Scholar 

  17. Bafetti, L., Young, N., Itoh, Y. & Stack, M. S. Intact vitronectin induces matrix metalloproteinase–2 and tissue inhibitor of metalloproteinase–2 expression and enhanced cellular invasion by melanoma cells. J. Biol. Chem. 273, 143–149 (1998).

    Article  CAS  Google Scholar 

  18. Talbot, D. C. & Brown, P. D. Experimental and clinical studies on the use of matrix metalloproteinase inhibitors for the treatment of cancer. Eur. J. Cancer 32, 2528– 2533 (1996).

    Article  Google Scholar 

  19. Narumiya, S. The small GTPase Rho: cellular functions and signal transduction. J. Biochem. (Tokyo) 120, 215–228 (1996).

    Article  CAS  Google Scholar 

  20. Madaule, P. et al. Role of citron kinase as a target of the small GTPase Rho in cytokinesis. Nature 394, 491– 494 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank T. Murozono for measuring the Y–27632 concentration. This work was supported in part by Grants–in–Aid for Cancer Research for a new 10–year strategy for cancer control from the Ministry of Health and Welfare of Japan, and for Specially Promoted Research from the Ministry of Education, Science, Sports, and Culture of Japan, as well as by grants from the Yamanouchi Foundation for Research on Metabolic Disease, the Uehara Memorial Foundation, the Naito Foundation, the Ichiro Kanahara Foundation (1997), and the Human Frontier Science Program.

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Correspondence to Kazuyuki Itoh.

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Itoh, K., Yoshioka, K., Akedo, H. et al. An essential part for Rho–associated kinase in the transcellular invasion of tumor cells. Nat Med 5, 221–225 (1999). https://doi.org/10.1038/5587

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