Integrin engagement differentially modulates epithelial cell motility by RhoA/ROCK and PAK1.
Integrin-ligand binding regulates tumor cell motility and invasion. Cell migration also involves the Rho GTPases that control the interplay between adhesion receptors and the cytoskeleton. We evaluated how specific extracellular matrix ligands modulate Rho GTPases and control motility of human squamous cell carcinoma cells. On laminin-5 substrates, the epithelial cells rapidly spread and migrated, but on type I collagen the cells spread slowly and showed reduced motility. We found that RhoA activity was suppressed in cells attached to laminin-5 through the alpha3 integrin receptor. In contrast, RhoA was strongly activated in cells bound to type I collagen and this was mediated by the alpha2 integrin. Inhibiting the RhoA pathway by expression of a dominant-negative RhoA mutant or by directly inhibiting ROCK, reduced focal adhesion formation and enhanced cell migration on type I collagen. Cdc42 and Rac and their downstream target PAK1 were activated following adhesion to laminin-5. PAK1 activation induced by laminin-5 was suppressed by expression of a dominant-negative Cdc42. Moreover, constitutively active PAK1 stimulated migration on collagen I substrates. Our results indicate that in squamous epithelial cells, collagen-alpha2beta1 integrin binding activates RhoA, slowing cell locomotion, whereas laminin-5-alpha3beta1 integrin interaction inhibits RhoA and activates PAK1, stimulating cell migration. The data demonstrate that specific ligand-integrin pairs regulate cell motility differentially by selectively modulating activities of Rho GTPases and their effectors.
Zhou
Hua
H
Department of Cell and Tissue Biology, University of California, San Francisco, California 94143, USA.
Kramer
Randall H
RH
D000256:Adenoviridae / Q000235:genetics; D002294:Carcinoma, Squamous Cell / Q000378:metabolism; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000737:chemistry*; D045744:Cell Line, Tumor; D002465:Cell Movement; D024042:Collagen Type I / Q000737:chemistry; D003599:Cytoskeleton / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology*; D005109:Extracellular Matrix / Q000378:metabolism; D005799:Genes, Dominant; D006801:Humans; D015151:Immunoblotting; D039422:Integrin alpha3 / Q000737:chemistry; D016023:Integrins / Q000737:chemistry*; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D008856:Microscopy, Fluorescence; D009154:Mutation; D010641:Phenotype; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D013997:Time Factors; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism; D054462:p21-Activated Kinases; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein / Q000737:chemistry*; D000097794:Kalinin
D015815:Cell Adhesion Molecules; D024042:Collagen Type I; D039422:Integrin alpha3; D016023:Integrins; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D017346:Protein Serine-Threonine Kinases; D020764:cdc42 GTP-Binding Protein; D054462:p21-Activated Kinases; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein; D000097794:Kalinin; C515903:PAK1 protein, human