title: p190RhoGEF (Rgnef) promotes colon carcinoma tumor progression via interaction with focal adhesion kinase.
abstract: Focal adhesion kinase (FAK) functions downstream of integrins and growth factor receptors to promote tumor cell motility and invasion. In colorectal cancer, FAK is activated by amidated gastrin, a protumorigenic hormone. However, it is unclear how FAK receives signals from the gastrin receptor or other G-protein-coupled receptors that can promote cell motility and invasion. The Rho guanine-nucleotide exchange factor p190RhoGEF (Rgnef) binds FAK and facilitates fibroblast focal adhesion formation on fibronectin. Here we report that Rgnef mRNA and protein expression are significantly increased during colorectal tumor progression. In human colon carcinoma cells, Rgnef forms a complex with FAK and upon gastrin stimulation, FAK translocates to newly-forming focal adhesions where it facilitates tyrosine phosphorylation of paxillin. short hairpin (shRNA)-mediated knockdown of Rgnef or FAK, or pharmacological inhibition of FAK activity, is sufficient to block gastrin-stimulated paxillin phosphorylation, cell motility, and invadopodia formation in a manner dependent upon upstream cholecystokinin-2 receptor expression. Overexpression of the C-terminal region of Rgnef (Rgnef-C, amino acid 1,279-1,582) but not Rgnef-CΔFAK (amino acid 1,302-1,582 lacking the FAK binding site) disrupted endogenous Rgnef-FAK interaction and prevented paxillin phosphorylation and cell motility stimulated by gastrin. Rgnef-C-expressing cells formed smaller, less invasive tumors with reduced tyrosine phosphorylation of paxillin upon orthotopic implantation, compared with Rgnef-CΔFAK-expressing cells. Our studies identify Rgnef as a novel regulator of colon carcinoma motility and invasion, and they show that a Rgnef-FAK linkage promotes colon carcinoma progression in vivo.
authors.affiliation: Department of Reproductive Medicine, Moores Cancer Center, UCSD, La Jolla, California 92093, USA.
authors.forename: Hong-Gang
authors.initials: HG
authors.lastname: Yu
authors.forename: Ju-Ock
authors.initials: JO
authors.lastname: Nam
authors.forename: Nichol L G
authors.initials: NL
authors.lastname: Miller
authors.forename: Isabelle
authors.initials: I
authors.lastname: Tanjoni
authors.forename: Colin
authors.initials: C
authors.lastname: Walsh
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.forename: Linda
authors.initials: L
authors.lastname: Kim
authors.forename: Xiao Lei
authors.initials: XL
authors.lastname: Chen
authors.forename: Alok
authors.initials: A
authors.lastname: Tomar
authors.forename: Ssang-Taek
authors.initials: ST
authors.lastname: Lim
authors.forename: David D
authors.initials: DD
authors.lastname: Schlaepfer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018938:Caco-2 Cells; D002465:Cell Movement / Q000502:physiology; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D018450:Disease Progression; D004789:Enzyme Activation; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D051417:Focal Adhesion Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005755:Gastrins / Q000378:metabolism; D055785:Gene Knockdown Techniques; D020662:Guanine Nucleotide Exchange Factors / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D045325:HCT116 Cells; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D051419:Paxillin / Q000378:metabolism; D010766:Phosphorylation; D015398:Signal Transduction
chemical_list: C551174:ARHGEF28 protein, human; D005755:Gastrins; D020662:Guanine Nucleotide Exchange Factors; C496061:PXN protein, human; D051419:Paxillin; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human