The Motogenic Effects of Cyclic Mechanical Strain on Intestinal Epithelial Monolayer Wound Closure Are Matrix Dependent

Jian-hu, Zhang; Owen, Cheri R.; Sanders, Matthew A.; Turner, Jerrold R.; Basson, Marc D.

doi:10.1053/j.gastro.2006.08.007

Cited by 47 publications

(106 citation statements)

References 78 publications

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“…The literature suggests that the extracellular matrix plays a vital role in regulating cell attachment, migration, and wound healing (36,37) In this context, a recent study (38) has elegantly shown that FN promotes epithelial cell migration/restitution in response to strain in Caco2 cells by inducing phosphorylation and activation of extracellular signal-regulated kinase signaling pathway and myosin light kinase. Our data demonstrate that FN induces NF-B activation that is required for attachment of cells to FN matrix.…”

Section: Discussionmentioning

confidence: 99%

Epithelial-derived Fibronectin Expression, Signaling, and Function in Intestinal Inflammation

Kolachala

Bajaj

Wang

et al. 2007

Journal of Biological Chemistry

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Fibronectin (FN) is a multifunctional extracellular matrix protein that plays an important role in cell proliferation, adhesion, and migration. FN expression or its role in colitis is not known. The goal of this study is to characterize FN expression, regulation, and role during intestinal inflammation. Wild-type and transgenic mice expressing luciferase under the control of the human FN promoter, given water or 3% dextran sodium sulfate, were used as animal models of colitis. The Caco2-BBE model intestinal epithelial cell line was used for in vitro studies. Mucosal tissue damage requiring efficient wound healing is a cardinal feature of inflammatory bowel disease as well as other intestinal inflammatory conditions, including radiation enteritis, chronic ischemic enteritis, and cystic fibrosis. Inappropriate or ineffective tissue repair underlies the persistence of disease symptoms and results in complications such as fibrosis or fistula formation. Epithelial response to injury is a complex process and cell-matrix interactions play important roles in the healing response. Extracellular matrix (ECM) 2 proteins such as fibronectin (FN), collagen, and laminin have been demonstrated to play a critical role in the wound healing process. In this study, we examined the expression and role of fibronectin in intestinal inflammation.FN is a high molecular weight adhesive glycoprotein that is found in basement membrane, lamina propria, and connective tissue matrices in the intestine (insoluble form) and in body fluids (soluble form). FN usually exists as a dimer composed of two nearly identical ϳ250-kDa subunits linked covalently near their C termini by a pair of disulfide bonds (1, 2). Each monomer is an extended and flexible molecule that is folded in a series of repeating protein domains known as type I, II, and III repeats. These domains are resistant to proteolysis and contain binding sites for other ECM proteins (e.g. collagen), cell-surface receptors (integrins), blood protein derivatives (fibrin), and glycosaminoglycans (heparin). There are more than 20 splice variants of FN. FN exists mainly in two forms, soluble plasma FN and less soluble cellular FN. Plasma FN is synthesized predominantly in the liver by hepatocytes and forms a soluble FN dimer as compared with tissue FN, which forms disulfide cross-linked fibrils and is deposited as a matrix.FN is widely expressed by multiple cell types and is critically important in vertebrate development, as demonstrated by the early embryonic lethality of mice with targeted inactivation of the FN gene (3). FN mediates a wide variety of cellular interactions with the ECM and plays important roles in cell adhesion, migration, growth, and differentiation (4, 5). Thus, not surprisingly, altered deposition of FN matrix has been correlated with several pathological states. For example, increased deposition of an FN-rich matrix has been associated with atherosclerosis and fibrosis, while restoration of FN matrix assembly in transformed cells has been correlated with a reduction in ...

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Section: Discussionmentioning

confidence: 99%

Epithelial-derived Fibronectin Expression, Signaling, and Function in Intestinal Inflammation

Kolachala

Bajaj

Wang

et al. 2007

Journal of Biological Chemistry

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“…Human intestinal epithelial Caco-2BBE cells, an established highly differentiated human colonocyte cell line (30,54), and nontransformed IEC-6 rat intestinal epithelial cells were maintained at 37°C in 5% CO2 as previously described (74).…”

Section: Methodsmentioning

confidence: 99%

“…Cells were plated on Flexwell plates and maintained in a 37°C humidified incubator with 5% CO 2. Once the cell monolayers were confluent, they were subjected to mechanical deformation using the Flexcell Strain Unit (FX-4000; Flexcell, McKeesport, PA) as described previously (74). Briefly, cells were subjected to cyclic deformation and relaxation at a magnitude of 10% strain and a frequency of 10 cycles/min by a computer-controlled 20-kPa vacuum.…”

Section: Methodsmentioning

confidence: 99%

“…Previous in vitro work from our laboratory suggests that cyclic strain stimulates Caco-2 and intestinal epithelial cell-6 (IEC-6) migration across a fibronectin matrix (74). This process requires the activation of Src, phosphatidylinositol 3-kinase (PI3K), and Akt, as well as a novel Src-independent phosphorylation of focal adhesion kinase (FAK) at Tyr925 (7,21).…”

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confidence: 99%

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ILK mediates the effects of strain on intestinal epithelial wound closure

Yuan

Sanders

Basson

2011

American Journal of Physiology-Cell Physiology

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The intestinal epithelium is subjected to repetitive deformation during normal gut function by peristalsis and villous motility. Such repetitive strain promotes intestinal epithelial migration across fibronectin in vitro, but signaling mediators for this are poorly understood. We hypothesized that integrin-linked kinase (ILK) mediates strain-stimulated migration in intestinal epithelial cells cultured on fibronectin. ILK kinase activity increased rapidly 5 min after strain induction in both Caco-2 and intestinal epithelial cell-6 (IEC-6) cells. Wound closure in response to strain was reduced in ILK small interfering RNA (siRNA)-transfected Caco-2 cell monolayers when compared with control siRNA-transfected Caco-2 cells. Pharmacological blockade of phosphatidylinositol-3 kinase (PI3K) or Src or reducing Src by siRNA prevented strain activation of ILK. ILK coimmunoprecipitated with focal adhesion kinase (FAK), and this association was decreased by mutation of FAK Tyr925 but not FAK Tyr397. Strain induction of FAK Tyr925 phosphorylation but not FAK Tyr397 or FAK Tyr576 phosphorylation was blocked in ILK siRNA-transfected cells. ILK-Src association was stimulated by strain and was blocked by the Src inhibitor PP2. Finally, ILK reduction by siRNA inhibited strain-induced phosphorylation of myosin light chain and Akt. These results suggest a strain-dependent signaling pathway in which ILK association with FAK and Src mediates the subsequent downstream strain-induced motogenic response and suggest that ILK induction by repetitive deformation may contribute to recovery from mucosal injury and restoration of the mucosal barrier in patients with prolonged ileus. ILK may therefore be an important target for intervention to maintain the mucosa in such patients.

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“…15 ERK activation is also noted to occur with the migration of epithelial cells on a fibronectin framework. 16 The enhanced migration observed in the Takagi et al study with the administration of ES could therefore reflect the establishment of a new extracellular matrix framework by the administration of ES through which the epithelial cells can migrate and not necessarily indicate a direct effect of ES on ERK activation, that is, the cells have the framework on which to migrate, and thus ERK is activated. As ERK is required for intestinal epithelial cell migration, 16 it was thus not surprising that the inhibition of ERK resulted in reduced cellular migration and the speed of wound healing independent of ES administration.…”

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confidence: 91%