Extracellular engagement of ADAM12 induces clusters of invadopodia with localized ectodomain shedding activity

Albrechtsen, Reidar; Stautz, Dorte; Sanjay, Archana; Kveiborg, Marie; Wewer, Ulla M.

doi:10.1016/j.yexcr.2010.10.003

Cited by 42 publications

(40 citation statements)

References 56 publications

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“…During cancer progression at primary and secondary sites, the tumor cells invariably interact with extracellular matrix (ECM) 2 components and alter the arrangement of cell-cell and cell-matrix adhesion (1).…”

Section: Together These Results Indicate That Ecti Inhibits the Invamentioning

confidence: 99%

“…Invadopodia, protrusions of the plasma membrane, are adhesive actin-rich structures located on the ventral cell surface of invasive tumor cells. In this manner, invadopodia establish focal contact with the substratum and play an intricate role in focal degradation of the extracellular matrix during cell invasion (2)(3)(4)(5). Active smooth muscle filamentous actin polymerization, induction of membrane curvature, rapid turnover of cell-matrix adhesions, and local modulation of contractile forces are all likely to play a central role in the promotion of invadopodium formation (6).…”

Section: Together These Results Indicate That Ecti Inhibits the Invamentioning

confidence: 99%

“…Several proteolytic enzymes are localized at the tip of invadopodia, including membrane type 1 metalloprotease (MT1-MMP) or MMP-14, secreted MMPs (MMP-2 and MMP-9), and a disintegrin and metalloproteinase-12 (2,20,21). To date the major contribution in cancer invasion attributed to MMPs is the degradation of the ECM barrier (1,22,23).…”

Section: Together These Results Indicate That Ecti Inhibits the Invamentioning

confidence: 99%

See 2 more Smart Citations

Enterolobium contortisiliquum Trypsin Inhibitor (EcTI), a Plant Proteinase Inhibitor, Decreases in Vitro Cell Adhesion and Invasion by Inhibition of Src Protein-Focal Adhesion Kinase (FAK) Signaling Pathways

Paula

Coulson‐Thomas

Ferreira

et al. 2012

Journal of Biological Chemistry

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Background:The effect of Enterolobium contortisiliquum trypsin inhibitor (EcTI) on the adhesion, migration, and invasion of gastric cancer cells. Results: EcTI inhibited adhesion, migration, and cell invasion and decreased Src-FAK signaling. Conclusion: EcTI inhibits the invasion of gastric cancer cells through alterations in integrin-dependent cell signaling pathways. Significance: Inhibition of invadopodia formation may be an attractive approach for cancer therapy.

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Section: Together These Results Indicate That Ecti Inhibits the Invamentioning

confidence: 99%

Section: Together These Results Indicate That Ecti Inhibits the Invamentioning

confidence: 99%

Section: Together These Results Indicate That Ecti Inhibits the Invamentioning

confidence: 99%

See 1 more Smart Citation

Enterolobium contortisiliquum Trypsin Inhibitor (EcTI), a Plant Proteinase Inhibitor, Decreases in Vitro Cell Adhesion and Invasion by Inhibition of Src Protein-Focal Adhesion Kinase (FAK) Signaling Pathways

Paula

Coulson‐Thomas

Ferreira

et al. 2012

Journal of Biological Chemistry

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show abstract

“…For example, in an experimental overexpression system, ADAM12, a protease which processes growth factor ligands and localizes to invadopodia, induces clusters of invadopodia where ectodomain shedding of EGFR ligands occurs. 70,71 This result suggests that invadopodia may provide spatio-temporal control of growth factors to control the invasive and metastatic potential of cancer cells in addition to their role of proteolyzing ECM.…”

Section: Growth Factorsmentioning

confidence: 99%

Regulation of invadopodia by the tumor microenvironment

Gould

Courtneidge

2014

Cell Adhesion & Migration

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“…Pioneered by Wen-Tien Chen in the early 1980's 4,14,15 , coating fluorescently labeled extracellular proteins on glass coverslips for subsequent microscopic analysis has emerged as the primary technique in evaluating invadopodia function across a wide range of cell types. The prescribed protocol demonstrates the basic method used for preparing gelatin-coated coverslips that form a collagenous layer less than 2 μm thick suitable for detection of extracellular matrix degradation by cells in most conventional fluorescent and confocal microscopes 11,[16][17][18] , similar to what has been previously described [19][20][21] . These properties allow for rapid production of coated coverslips capable of detecting the initial onset of matrix degradation.…”

Section: Discussionmentioning

confidence: 94%

Quantitative Measurement of Invadopodia-mediated Extracellular Matrix Proteolysis in Single and Multicellular Contexts

Martin

Hayes

Walk

et al. 2012

JoVE

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Cellular invasion into local tissues is a process important in development and homeostasis. Malregulated invasion and subsequent cell movement is characteristic of multiple pathological processes, including inflammation, cardiovascular disease and tumor cell metastasis 1 . Focalized proteolytic degradation of extracellular matrix (ECM) components in the epithelial or endothelial basement membrane is a critical step in initiating cellular invasion. In tumor cells, extensive in vitro analysis has determined that ECM degradation is accomplished by ventral actin-rich membrane protrusive structures termed invadopodia 2,3 . Invadopodia form in close apposition to the ECM, where they moderate ECM breakdown through the action of matrix metalloproteinases (MMPs). The ability of tumor cells to form invadopodia directly correlates with the ability to invade into local stroma and associated vascular components 3 .Visualization of invadopodia-mediated ECM degradation of cells by fluorescent microscopy using dye-labeled matrix proteins coated onto glass coverslips has emerged as the most prevalent technique for evaluating the degree of matrix proteolysis and cellular invasive potential 4,5 . Here we describe a version of the standard method for generating fluorescently-labeled glass coverslips utilizing a commercially available Oregon Green-488 gelatin conjugate. This method is easily scaled to rapidly produce large numbers of coated coverslips. We show some of the common microscopic artifacts that are often encountered during this procedure and how these can be avoided. Finally, we describe standardized methods using readily available computer software to allow quantification of labeled gelatin matrix degradation mediated by individual cells and by entire cellular populations. The described procedures provide the ability to accurately and reproducibly monitor invadopodia activity, and can also serve as a platform for evaluating the efficacy of modulating protein expression or testing of anti-invasive compounds on extracellular matrix degradation in single and multicellular settings. Video LinkThe video component of this article can be found at http://www.jove.com/video/4119/ Protocol Production of Oregon Green 488-gelatin Coated Coverslips1. Prepare an unlabeled 5% (w/w) stock gelatin/sucrose solution by adding 1.25 g gelatin and 1.25 g sucrose in PBS to a final volume of 50 ml.Warm the stock gelatin solution to 37 °C and ensure it is entirely melted before use. Store the final mixture at 4 °C. 2. Clean 13 mm diameter #1 glass coverslips by placing an individual coverslip into each well of a 24 well plastic tissue culture plate. Add 500 μl of 20% nitric acid to each well and incubate for 30 min. Aspirate the nitric acid solution and wash coverslips three times with deionized water. 3. Coat coverslips with 500 μl of 50 μg/ml poly-L-lysine (prepared from 0.1% stock solution and diluted in deionized water) to each well for 20 min at room temperature. Aspirate the solution and wash three times with PBS. Poly-L-lysine coa...

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Extracellular engagement of ADAM12 induces clusters of invadopodia with localized ectodomain shedding activity

Cited by 42 publications

References 56 publications

Enterolobium contortisiliquum Trypsin Inhibitor (EcTI), a Plant Proteinase Inhibitor, Decreases in Vitro Cell Adhesion and Invasion by Inhibition of Src Protein-Focal Adhesion Kinase (FAK) Signaling Pathways

Enterolobium contortisiliquum Trypsin Inhibitor (EcTI), a Plant Proteinase Inhibitor, Decreases in Vitro Cell Adhesion and Invasion by Inhibition of Src Protein-Focal Adhesion Kinase (FAK) Signaling Pathways

Regulation of invadopodia by the tumor microenvironment

Quantitative Measurement of Invadopodia-mediated Extracellular Matrix Proteolysis in Single and Multicellular Contexts

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