BACKGROUND: Although there is extensive evidence for the amoeboid invasiveness of cancer cells in vitro, much less is known about the role of amoeboid invasiveness in metastasis and the importance of Rho/ROCK/MLC signaling in this process. RESULTS: We analyzed the dependence of amoeboid invasiveness of rat and chicken sarcoma cells and the metastatic activity of chicken cells on individual elements of the Rho/ROCK/MLC pathway. In both animal models, inhibition of Rho, ROCK or MLC resulted in greatly decreased cell invasiveness in vitro, while inhibition of extracellular proteases using a broad spectrum inhibitor did not have a significant effect. The inhibition of both Rho activity and MLC phosphorylation by dominant negative mutants led to a decreased capability of chicken sarcoma cells to metastasize. Moreover, the overexpression of RhoA in non-metastatic chicken cells resulted in the rescue of both invasiveness and metastatic capability. Rho and ROCK, unlike MLC, appeared to be directly involved in the maintenance of the amoeboid phenotype, as their inhibition resulted in the amoeboid-mesenchymal transition in analyzed cell lines. CONCLUSION: Taken together, these results suggest that protease-independent invasion controlled by elements of the Rho/ROCK/MLC pathway can be frequently exploited by metastatic sarcoma cells.

Department of Cell Biology Faculty of Science Charles University Prague Viničná 7 12843 Prague 2 Czech Republic

Zobrazit více v PubMed

Steeg PS, Theodorescu D. Metastasis: a therapeutic target for cancer. Nat Clin Pract Oncol. 2008;5:206–219. doi: 10.1038/ncponc1066. PubMed DOI PMC

Deryugina EI, Bourdon MA. Tenascin mediates human glioma cell migration and modulates cell migration on fibronectin. J Cell Sci. 1996;109(Pt 3):643–652. PubMed

Pollard TD, Borisy GG. Cellular motility driven by assembly and disassembly of actin filaments. Cell. 2003;112:453–465. doi: 10.1016/S0092-8674(03)00120-X. PubMed DOI

Yamaguchi H, Wyckoff J, Condeelis J. Cell migration in tumors. Curr Opin Cell Biol. 2005;17:559–564. doi: 10.1016/j.ceb.2005.08.002. PubMed DOI

Mierke CT, Rosel D, Fabry B, Brabek J. Contractile forces in tumor cell migration. Eur J Cell Biol. 2008;87:669–676. doi: 10.1016/j.ejcb.2008.01.002. PubMed DOI PMC

Hall A. Rho GTPases and the actin cytoskeleton. Science. 1998;279:509–514. doi: 10.1126/science.279.5350.509. PubMed DOI

Ridley AJ. Rho GTPases and cell migration. J Cell Sci. 2001;114:2713–2722. PubMed

Wolf K, Muller R, Borgmann S, Brocker EB, Friedl P. Amoeboid shape change and contact guidance: T-lymphocyte crawling through fibrillar collagen is independent of matrix remodeling by MMPs and other proteases. Blood. 2003;102:3262–3269. doi: 10.1182/blood-2002-12-3791. PubMed DOI

Sahai E, Marshall CJ. Differing modes of tumour cell invasion have distinct requirements for Rho/ROCK signalling and extracellular proteolysis. Nat Cell Biol. 2003;5:711–719. doi: 10.1038/ncb1019. PubMed DOI

Polyak K, Weinberg RA. Transitions between epithelial and mesenchymal states: acquisition of malignant and stem cell traits. Nat Rev Cancer. 2009;9:265–273. doi: 10.1038/nrc2620. PubMed DOI

Rosel D, Brabek J, Tolde O, Mierke CT, Zitterbart DP, Raupach C, Bicanova K, Kollmannsberger P, Pankova D, Vesely P. et al.Up-regulation of Rho/ROCK signaling in sarcoma cells drives invasion and increased generation of protrusive forces. Mol Cancer Res: MCR. 2008;6:1410–1420. doi: 10.1158/1541-7786.MCR-07-2174. PubMed DOI

Wyckoff JB, Pinner SE, Gschmeissner S, Condeelis JS, Sahai E. ROCK- and myosin-dependent matrix deformation enables protease-independent tumor-cell invasion in vivo. Curr Biol: CB. 2006;16:1515–1523. doi: 10.1016/j.cub.2006.05.065. PubMed DOI

Jay PY, Pham PA, Wong SA, Elson EL. A mechanical function of myosin II in cell motility. J Cell Sci. 1995;108(Pt 1):387–393. PubMed

Lauffenburger DA, Horwitz AF. Cell migration: a physically integrated molecular process. Cell. 1996;84:359–369. doi: 10.1016/S0092-8674(00)81280-5. PubMed DOI

Friedl P, Wolf K. Plasticity of cell migration: a multiscale tuning model. J Cell Biol. 2010;188:11–19. doi: 10.1083/jcb.200909003. PubMed DOI PMC

Sahai E. Illuminating the metastatic process. Nat Rev Cancer. 2007;7:737–749. PubMed

Pankova K, Rosel D, Novotny M, Brabek J. The molecular mechanisms of transition between mesenchymal and amoeboid invasiveness in tumor cells. Cell Mol Life Sci: CMLS. 2010;67:63–71. doi: 10.1007/s00018-009-0132-1. PubMed DOI PMC

Tolde O, Rosel D, Vesely P, Folk P, Brabek J. The structure of invadopodia in a complex 3D environment. Eur J Cell Biol. 2010;89:674–680. doi: 10.1016/j.ejcb.2010.04.003. PubMed DOI

Tolde O, Rosel D, Janostiak R, Vesely P, Brabek J. Dynamics and morphology of focal adhesions in complex 3D environment. Folia biologica. 2012;58:177–184. PubMed

Cermak V, Kosla J, Plachy J, Trejbalova K, Hejnar J, Dvorak M. The transcription factor EGR1 regulates metastatic potential of v-src transformed sarcoma cells. Cell Mol Life Sci: CMLS. 2010;67:3557–3568. doi: 10.1007/s00018-010-0395-6. PubMed DOI PMC

Sabeh F, Shimizu-Hirota R, Weiss SJ. Protease-dependent versus -independent cancer cell invasion programs: three-dimensional amoeboid movement revisited. J Cell Biol. 2009;185:11–19. doi: 10.1083/jcb.200807195. PubMed DOI PMC

Micuda S, Rosel D, Ryska A, Brabek J. ROCK inhibitors as emerging therapeutic candidates for sarcomas. Curr Cancer Drug Tar. 2010;10:127–134. doi: 10.2174/156800910791054202. PubMed DOI

Ying H, Biroc SL, Li WW, Alicke B, Xuan JA, Pagila R, Ohashi Y, Okada T, Kamata Y, Dinter H. The Rho kinase inhibitor fasudil inhibits tumor progression in human and rat tumor models. Mol Cancer Ther. 2006;5:2158–2164. doi: 10.1158/1535-7163.MCT-05-0440. PubMed DOI

Xue F, Takahara T, Yata Y, Xia Q, Nonome K, Shinno E, Kanayama M, Takahara S, Sugiyama T. Blockade of Rho/Rho-associated coiled coil-forming kinase signaling can prevent progression of hepatocellular carcinoma in matrix metalloproteinase-dependent manner. Hepatol Res: the official journal of the Japan Society of Hepatology. 2008;38:810–817. doi: 10.1111/j.1872-034X.2008.00333.x. PubMed DOI

Hager MH, Morley S, Bielenberg DR, Gao S, Morello M, Holcomb IN, Liu W, Mouneimne G, Demichelis F, Kim J. et al.DIAPH3 governs the cellular transition to the amoeboid tumour phenotype. EMBO Mol Med. 2012;4:743–760. doi: 10.1002/emmm.201200242. PubMed DOI PMC

Belletti B, Nicoloso MS, Schiappacassi M, Berton S, Lovat F, Wolf K, Canzonieri V, D'Andrea S, Zucchetto A, Friedl P. et al.Stathmin activity influences sarcoma cell shape, motility, and metastatic potential. Mol Biol Cell. 2008;19:2003–2013. doi: 10.1091/mbc.E07-09-0894. PubMed DOI PMC

Parri M, Taddei ML, Bianchini F, Calorini L, Chiarugi P. EphA2 reexpression prompts invasion of melanoma cells shifting from mesenchymal to amoeboid-like motility style. Cancer Res. 2009;69:2072–2081. PubMed

Taddei ML, Parri M, Angelucci A, Bianchini F, Marconi C, Giannoni E, Raugei G, Bologna M, Calorini L, Chiarugi P. EphA2 induces metastatic growth regulating amoeboid motility and clonogenic potential in prostate carcinoma cells. Mol Cancer Res: MCR. 2011;9:149–160. doi: 10.1158/1541-7786.MCR-10-0298. PubMed DOI

Belgiovine C, Frapolli R, Bonezzi K, Chiodi I, Favero F, Mello-Grand M, Dei Tos AP, Giulotto E, Taraboletti G, D'Incalci M, Mondello C. Reduced expression of the ROCK inhibitor Rnd3 is associated with increased invasiveness and metastatic potential in mesenchymal tumor cells. PLoS One. 2010;5:e14154. doi: 10.1371/journal.pone.0014154. PubMed DOI PMC

Cavanna T, Pokorna E, Vesely P, Gray C, Zicha D. Evidence for protein 4.1B acting as a metastasis suppressor. J Cell Sci. 2007;120:606–616. doi: 10.1242/jcs.000273. PubMed DOI

Vesely P, Donner L, Cinatl J, Sovova V. Interaction of Rous sarcoma virus with rat embryo fibroblasts of inbred Lewis strain in vitro. Folia biologica. 1968;14:457–465. PubMed

Conrad S, Genth H, Hofmann F, Just I, Skutella T. Neogenin-RGMa signaling at the growth cone is bone morphogenetic protein-independent and involves RhoA, ROCK, and PKC. J Biol Chem. 2007;282:16423–16433. doi: 10.1074/jbc.M610901200. PubMed DOI

Croft DR, Coleman ML, Li S, Robertson D, Sullivan T, Stewart CL, Olson MF. Actin-myosin-based contraction is responsible for apoptotic nuclear disintegration. J Cell Biol. 2005;168:245–255. doi: 10.1083/jcb.200409049. PubMed DOI PMC

Janostiak R, Tolde O, Bruhova Z, Novotny M, Hanks SK, Rosel D, Brabek J. Tyrosine phosphorylation within the SH3 domain regulates CAS subcellular localization, cell migration, and invasiveness. Mol Biol Cell. 2011;22:4256–4267. doi: 10.1091/mbc.E11-03-0207. PubMed DOI PMC

Plachy J, Vilhelmova M. Syngeneic lines of chickens. VII. The lines derived from the recombinants at the B complex (MHC) of Rous sarcoma regressor and progressor inbred lines of chickens. Folia biologica. 1984;30:189–201. PubMed

Suppression of the growth and metastasis of mouse melanoma by Taenia crassiceps and Mesocestoides corti tapeworms

The emerging role of microtubules in invasion plasticity

RNA-seq Characterization of Melanoma Phenotype Switch in 3D Collagen after p38 MAPK Inhibitor Treatment

High-throughput transcriptomic and proteomic profiling of mesenchymal-amoeboid transition in 3D collagen

Increased Level of Long Non-Coding RNA MALAT1 is a Common Feature of Amoeboid Invasion

Migrastatics-Anti-metastatic and Anti-invasion Drugs: Promises and Challenges