title: Role of RhoA and its effectors ROCK and mDia1 in the modulation of deformation-induced FAK, ERK, p38, and MLC motogenic signals in human Caco-2 intestinal epithelial cells.
abstract: Repetitive deformation enhances intestinal epithelial migration across tissue fibronectin. We evaluated the contribution of RhoA and its effectors Rho-associated kinase (ROK/ROCK) and mammalian diaphanous formins (mDia1) to deformation-induced intestinal epithelial motility across fibronectin and the responsible focal adhesion kinase (FAK), extracellular signal-regulated kinase (ERK), p38, and myosin light chain (MLC) signaling. We reduced RhoA, ROCK1, ROCK2, and mDia1 by smart-pool double-stranded short-interfering RNAs (siRNA) and pharmacologically inhibited RhoA, ROCK, and FAK in human Caco-2 intestinal epithelial monolayers on fibronectin-coated membranes subjected to 10% repetitive deformation at 10 cycles/min. Migration was measured by wound closure. Stimulation of migration by deformation was prevented by exoenzyme C3, Y27632, or selective RhoA, ROCK1, and ROCK2 or mDia1 siRNAs. RhoA, ROCK inhibition, or RhoA, ROCK1, ROCK2, mDia1, and FAK reduction by siRNA blocked deformation-induced nuclear ERK phosphorylation without preventing ERK phosphorylation in the cytoplasmic protein fraction. Furthermore, RhoA, ROCK inhibition or RhoA, ROCK1, ROCK2, and mDia1 reduction by siRNA also blocked strain-induced FAK-Tyr(925), p38, and MLC phosphorylation. These results suggest that RhoA, ROCK, mDia1, FAK, ERK, p38, and MLC all mediate the stimulation of intestinal epithelial migration by repetitive deformation. This pathway may be an important target for interventions to promote mechanotransduced mucosal healing during inflammation.
authors.affiliation: Department of Surgery, Michigan State University, Lansing, MI 48912, USA.
authors.forename: Lakshmi S
authors.initials: LS
authors.lastname: Chaturvedi
authors.forename: Harold M
authors.initials: HM
authors.lastname: Marsh
authors.forename: Marc D
authors.initials: MD
authors.lastname: Basson
mesh_terms: D036002:ADP Ribose Transferases / Q000494:pharmacology; D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000577:Amides / Q000494:pharmacology; D001905:Botulinum Toxins / Q000494:pharmacology; D018938:Caco-2 Cells; D002465:Cell Movement / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D000080865:Formins; D006801:Humans; D020935:MAP Kinase Signaling System; D040542:Mechanotransduction, Cellular; D018994:Myosin Light Chains / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000378:metabolism; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000577:Amides; C108962:DIAPH1 protein, human; D004791:Enzyme Inhibitors; D005353:Fibronectins; D000080865:Formins; D018994:Myosin Light Chains; D011725:Pyridines; D034741:RNA, Small Interfering; C066737:RHOA protein, human; C108830:Y 27632; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D051416:Focal Adhesion Protein-Tyrosine Kinases; C515851:ROCK1 protein, human; C515852:ROCK2 protein, human; D054460:rho-Associated Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D001905:Botulinum Toxins; D020742:rhoA GTP-Binding Protein