Dataset Information

The diacylglycerol kinase ?/atypical PKC/?1 integrin pathway in SDF-1? mammary carcinoma invasiveness.

ABSTRACT: Diacylglycerol kinase ? (DGK?), by phosphorylating diacylglycerol into phosphatidic acid, provides a key signal driving cell migration and matrix invasion. We previously demonstrated that in epithelial cells activation of DGK? activity promotes cytoskeletal remodeling and matrix invasion by recruiting atypical PKC at ruffling sites and by promoting RCP-mediated recycling of ?5?1 integrin to the tip of pseudopods. In here we investigate the signaling pathway by which DGK? mediates SDF-1?-induced matrix invasion of MDA-MB-231 invasive breast carcinoma cells. Indeed we showed that, following SDF-1? stimulation, DGK? is activated and localized at cell protrusion, thus promoting their elongation and mediating SDF-1? induced MMP-9 metalloproteinase secretion and matrix invasion. Phosphatidic acid generated by DGK? promotes localization at cell protrusions of atypical PKCs which play an essential role downstream of DGK? by promoting Rac-mediated protrusion elongation and localized recruitment of ?1 integrin and MMP-9. We finally demonstrate that activation of DGK?, atypical PKCs signaling and ?1 integrin are all essential for MDA-MB-231 invasiveness. These data indicates the existence of a SDF-1? induced DGK? - atypical PKC - ?1 integrin signaling pathway, which is essential for matrix invasion of carcinoma cells.

SUBMITTER: Rainero E

PROVIDER: S-EPMC4041662 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Publications

The diacylglycerol kinase α/atypical PKC/β1 integrin pathway in SDF-1α mammary carcinoma invasiveness.

Rainero Elena E Cianflone Cristina C Porporato Paolo Ettore PE Chianale Federica F Malacarne Valeria V Bettio Valentina V Ruffo Elisa E Ferrara Michele M Benecchia Fabio F Capello Daniela D Paster Wolfgang W Locatelli Irene I Bertoni Alessandra A Filigheddu Nicoletta N Sinigaglia Fabiola F Norman Jim C JC Baldanzi Gianluca G Graziani Andrea A

PloS one 20140602 6

Diacylglycerol kinase α (DGKα), by phosphorylating diacylglycerol into phosphatidic acid, provides a key signal driving cell migration and matrix invasion. We previously demonstrated that in epithelial cells activation of DGKα activity promotes cytoskeletal remodeling and matrix invasion by recruiting atypical PKC at ruffling sites and by promoting RCP-mediated recycling of α5β1 integrin to the tip of pseudopods. In here we investigate the signaling pathway by which DGKα mediates SDF-1α-induced ...[more]

PMID: 24887021

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Project description:BackgroundThe blood-spinal cord barrier (BSCB) is composed of a monolayer of endothelium linked with tight junctions and extracellular matrix (ECM)-rich basement membranes and is surrounded by astrocyte foot processes. Endothelial permeability is regulated by interaction between endothelial cells and ECM proteins. Fibronectin (FN) is a principal ECM component of microvessels. Excessive FN deposition disrupts cell-cell adhesion in fibroblasts through β1 integrin ligation. To determine whether excessive FN deposition contributes to the disruption of endothelial integrity, we used an in vitro model of the endothelial monolayer to investigate whether the FN inhibitor pUR4 prevents FN deposition into the subendothelial matrix and attenuates endothelial leakage.MethodsTo correlate the effects of excessive FN accumulation in microvessels on BSCB disruption, spinal nerve ligation-which induces BSCB leakage-was applied, and FN expression in the spinal cord was evaluated through immunohistochemistry and immunoblotting. To elucidate the effects by which pUR4 modulates endothelial permeability, brain-derived endothelial (bEND.3) cells treated with tumor necrosis factor (TNF)-α were used to mimic a leaky BSCB. A bEND.3 monolayer was preincubated with pUR4 before TNF-α treatment. The transendothelial electrical resistance (TEER) measurement and transendothelial permeability assay were applied to assess the endothelial integrity of the bEND.3 monolayer. Immunofluorescence analysis and immunoblotting were performed to evaluate the inhibitory effects of pUR4 on TNF-α-induced FN deposition. To determine the mechanisms underlying pUR4-mediated endothelial permeability, cell morphology, stress fiber formation, myosin light chain (MLC) phosphorylation, and β1 integrin-mediated signaling were evaluated through immunofluorescence analysis and immunoblotting.ResultsExcessive FN was accumulated in the microvessels of the spinal cord after spinal nerve ligation; moreover, pUR4 inhibited TNF-α-induced FN deposition in the bEND.3 monolayer and maintained intact TEER and endothelial permeability. Furthermore, pUR4 reduced cell morphology alteration, actin stress fiber formation, and MLC phosphorylation, thereby attenuating paracellular gap formation. Moreover, pUR4 reduced β1 integrin activation and downstream signaling.ConclusionspUR4 reduces TNF-α-induced β1 integrin activation by depleting ECM FN, leading to a decrease in endothelial hyperpermeability and maintenance of monolayer integrity. These findings suggest therapeutic benefits of pUR4 in pathological vascular leakage treatment.

Dataset Information

The diacylglycerol kinase ?/atypical PKC/?1 integrin pathway in SDF-1? mammary carcinoma invasiveness.

Publications

The diacylglycerol kinase α/atypical PKC/β1 integrin pathway in SDF-1α mammary carcinoma invasiveness.

Similar Datasets

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