We have previously shown that overexpression of focal adhesion kinase (FAK) in Chinese hamster ovary (CHO) cells promoted their migration on fibronectin. This effect was dependent the phosphorylation FAK at Tyr-397. residue known to serve as a binding site for both Src and phosphatidylinositol 3-kinase (PI3K), implying either one or are required promote cell migration. In this study, we examined role PI3K FAK-promoted demonstrated inhibitors, wortmannin LY294002, were able inhibit...

The Boyden chamber assay, originally introduced by for the analysis of leukocyte chemotaxis, is based on a two medium-filled compartments separated microporous membrane. In general, cells are placed in upper compartment and allowed to migrate through pores membrane into lower compartment, which chemotactic agents present. After an appropriate incubation time, between fixed stained, number that have migrated side determined. Therefore, chamber-based cell migration assay has also been called...

Fibroblasts derived from focal adhesion kinase (FAK)-null mouse embryos have a reduced migration rate and an increase in the number size of peripherally localized adhesions (Ilic, D., Furuta, Y., Kanazawa, S., Takeda, N., Sobue, K., Nakatsuji, Nomura, Fujimoto, J., Okada, M., Yamamoto, T. (1995)Nature 377, 539–544). In this study, we found that Y27632, specific inhibitor for Rho-associated (Rho-kinase), dramatically reversed round cell morphology FAK−/−cells to spread fibroblast-like shape...

Focal adhesion kinase (FAK) has been implicated to be a point of convergence integrin and growth factor signaling pathways. Here we report that FAK directly interacts with the hepatocyte receptor c-Met. Phosphorylation c-Met at Tyr-1349 and, lesser extent, Tyr-1356 is required for its interaction band 4.1 ezrin/radixin/moesin homology domain (FERM domain) FAK. The F2 subdomain FERM alone sufficient Met binding, in which patch basic residues (216KAKTLRK222) are critical interaction. Met-FAK...

Focal adhesion kinase (FAK) has been implicated to play a role in suppression of apoptosis. In this study, we have demonstrated that UV irradiation induced cleavage FAK and two its interacting proteins Src p130^Cas Madin-Darby canine kidney cells, concomitant with an increase cell death. The these upon was completely inhibited by ZVAD-FMK, broad range inhibitor caspases, apparently delayed Bcl2 overexpression. To examine if plays suppressing UV-induced apoptosis, stable lines...

Diabetic nephropathy progressed to end-stage renal disease (ESRD) is found in type 1 or 2 diabetes. Oxidative stress one of the precipitation factors diabetic nephropathy. Previously, Hibiscus sabdariffa Linnaeus and its polyphenol extracts were possess antioxidative effects. This study aimed investigate effect L. extract (HPE) streptozotocin (STZ) induced The results show that HPE reduced kidney mass by STZ significantly, as well improving hydropic change proximal convoluted tubules rats....

Podosomes are dynamic actin-enriched membrane structures that play an important role in invasive cell motility and extracellular matrix degradation. They often found to assemble into large rosettelike highly cells. However, the mechanism of this assembly remains obscure. In study, we identified focal adhesion kinase (FAK) as a key molecule necessary for assembly. Moreover, phosphorylation p130Cas suppression Rho signaling by FAK were be induce podosome rosettes. Finally, vimentin...

Mitotic spindles are microtubule-based structures, but increasing evidence indicates that filamentous actin (F-actin) and F-actin–based motors components of these structures. ADD1 (adducin-1) is an actin-binding protein has been shown to play important roles in the stabilization membrane cortical cytoskeleton cell–cell adhesions. In this study, we show associates with mitotic crucial for proper spindle assembly progression. Phosphorylation at Ser12 Ser355 by cyclin-dependent kinase 1 enables...

Vimentin intermediate filaments (VIFs), expressed in most mesenchymal and cancer cells, undergo dramatic reorganization during cell migration; however, the mechanism remains obscure. This study demonstrates that upon growth-factor stimulation, Src directly phosphorylates vimentin at Tyr117, leading to VIF disassembly into squiggles particles edge lamellipodia formation. The protein tyrosine phosphatase SHP2 counteracted effects on phosphorylation organization. VIFs formed by Y117D mutant...

Focal adhesion kinase (FAK) has been implicated to play a critical role in integrin-mediated control of cell behavior. However, it is unclear whether FAK also participates the regulation growth factor-elicited cellular functions. In this study, we have demonstrated that although overexpression Madin-Dardy canine kidney cells did not alter their property or ability form tubules within collagen gel upon hepatocyte factor (HGF) stimulation, apparently enhanced HGF-induced scattering. This...

Cell–matrix interactions play critical roles in a variety of physiological and pathological processes by regulating cell migration, proliferation, differentiation. Integrin family cell-adhesion receptors have been identified as the major for various extracellular matrix proteins shown to be capable transducing biochemical signals across plasma membrane regulate cellular functions. Protein tyrosine phosphorylation has implicated central role integrin-initiated signal transduction. Other...

We examined gene expression of hepatocyte growth factor (HGF) and HGF receptor (HGFR), or product proto-oncogene c-met (c-met), in smokers nonsmokers with adenocarcinoma (ADC) by suppression subtractive hybridization microarray techniques. Expression was confirmed RT-PCR. content the respective tumor mass nontumor lung tissue measured ELISA. pathologic samples localized immunohistochemistry situ hybridization. Our results indicate that overexpression HGFR frequently detected ADC cells,...

Diabetic nephropathy is characterized as the progressive development of renal insufficiency in a setting hyperglycemia. Previous studies indicate that reactive oxygen species (ROS) play an important role high glucose-induced injury. Cilostazol was reported to lower production superoxide significantly situ. We hypothesized cilostazol administration streptozotocin-induced diabetic rats exerts effects via improving oxidative stress. Male Sprague–Dawley were fed with (5 mg/kg or 25 mg/kg) for 12...

Protein kinase Cδ (PKCδ) has been implicated to play a crucial role in cell proliferation, differentiation and apoptosis. In this study, we have investigated the of PKCδ motility using Madin-Darby canine kidney cells. Overexpression promoted membrane protrusions, concomitant with increased motility. By contrast, suppression expression by RNA interference inhibited Moreover, fraction was detected at edge protrusions which it colocalized adducin, skeletal protein whose phosphorylation state is...

The clinical significance of STIM proteins and Orai Ca2+ channels in tumor progression has been demonstrated different types cancers. Podosomes are dynamic actin-rich cellular protrusions that facilitate cancer cell invasiveness by degrading extracellular matrix. Whether STIM1-dependent signaling facilitates invasion through affecting podosome formation remains unclear. Here we show the invasive fronts tissues overexpress STIM1, accompanied active store-operated entry (SOCE). Interfering...

Podosomes are dynamic actin-based membrane protrusions that important for extracellular matrix degradation and invasive cell motility. Individual podosomes often found to organize into large rosette-like structures in some types of cells, such as osteoclasts, endothelial Src-transformed fibroblasts, certain highly cancer cells. In this study, we show new podosome rosettes arise through one two mechanisms; de novo assembly or fission a pre-existing rosette fibroblasts. Fission is more...

Epithelial cells usually trigger their “migratory machinery” upon loss of adhesion to neighbors. This default is important for both physiological (e.g., wound healing) and pathological tumor metastasis) processes. However, the underlying mechanism such a remains unclear. In this study, we used human head neck squamous cell carcinoma (HNSCC) SAS as model found that cell–cell induced reactive oxygen species (ROS) generation vimentin expression, which were required migration adhesion. We...

Grb2-associated binder 1 (Gab1) is known to play an important role in hepatocyte growth factor (HGF) signaling, which rapidly becomes tyrosine-phosphorylated upon HGF stimulation. In this study, we found that the tyrosine phosphorylation of Gab1 cells derived from Src/Yes/Fyn null mouse embryos was ∼40% lower than their wild type counterparts Increased expression wild-type Src enhanced HGF-induced Gab1, and, contrast, kinase-deficient mutant or treatment specific inhibitor PP1 suppressed it....

The F-actin binding protein adducin plays an important role in plasma membrane stability, cell motility and cell-cell junctions. In this study, we demonstrate that α-adducin is mainly localized the nucleus of sparsely cultured epithelial cells, whereas it at junctions when cells are grown to confluence. Disruption adhesions induces a nuclear translocation α-adducin. Conversely, redistributed cytoplasm process establishing adhesions. We identify contains bipartite localization signal (NLS)...

Proper control of cell-cell adhesion is crucial for embryogenesis and tissue homeostasis. In this study, we show that protein kinase C (PKC)δ, a member the novel PKC subfamily, localizes at contacts epithelial cells through its C2-like domain in an F-actin-dependent manner. Upon hepatocyte growth factor stimulation, PKCδ phosphorylated activated by Src, which then phosphorylates E-cadherin Thr790. Phosphorylation Thr790 diminishes interaction with β-catenin impairs homophilic between...

Nuclear deformation has been observed in some cancer cells for decades, but its underlying mechanism and biological significance remain elusive. To address these questions, we employed human lung A549 cell line as a model context with transforming growth factor β (TGFβ)-induced epithelial-mesenchymal transition. Here, report that nuclear induced by TGFβ is concomitant increased phosphorylation of lamin A at Ser390, defective lamina genome instability. AKT2 Smad3 serve the downstream...

Protein kinase C (PKC) delta, a member of the novel PKC subfamily, has been shown to have an important role in cell proliferation, differentiation, apoptosis and motility. In this study, we investigated effect green fluorescent protein (GFP)-PKCdelta GFP-PKCalpha on cell-cell junctions Madin-Darby canine kidney (MDCK) cells found that only GFP-PKCdelta suppressed homophilic interactions between ectodomains E-cadherins, accompanied by weaker adhesion. The kinase-deficient mutant retained its...