scholarly journals Oncogenic H-Ras Reprograms Madin-Darby Canine Kidney (MDCK) Cell-derived Exosomal Proteins Following Epithelial-Mesenchymal Transition

2013 ◽  
Vol 12 (8) ◽  
pp. 2148-2159 ◽  
Author(s):  
Bow J. Tauro ◽  
Rommel A. Mathias ◽  
David W. Greening ◽  
Shashi K. Gopal ◽  
Hong Ji ◽  
...  
Keyword(s):  
Mdck Cell ◽  
Epithelial Mesenchymal Transition ◽  
Madin Darby Canine Kidney ◽  
Mesenchymal Transition ◽  
Darby Canine Kidney ◽  
Canine Kidney

scholarly journals STAT1 Is Required for Redifferentiation during Madin-Darby Canine Kidney Tubulogenesis

2010 ◽  
Vol 21 (22) ◽  
pp. 3926-3933 ◽  
Author(s):  
Minji Kim ◽  
Lucy Erin O'Brien ◽  
Sang-Ho Kwon ◽  
Keith E. Mostov
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Mdck Cells ◽  
Madin Darby Canine Kidney ◽  
Tubule Formation ◽  
Mesenchymal Transition ◽  
Phase Loss ◽  
Darby Canine Kidney ◽  
Canine Kidney ◽  
Constitutively Active

Tubule formation in vitro using Madin-Darby canine kidney (MDCK) epithelial cells consists mainly of two processes. First, the cells undergo a partial epithelial–mesenchymal transition (pEMT), losing polarity and migrating. Second, the cells redifferentiate, forming cords and then tubules with continuous lumens. We have shown previously that extracellular signal-regulated kinase activation is required for pEMT. However, the mechanism of how the pEMT phase is turned off and the redifferentiation phase is initiated is largely unknown. To address the central question of the sequential control of these two phases, we used MDCK cells grown as cysts and treated with hepatocyte growth factor to model tubulogenesis. We show that signal transducer and activator of transcription (STAT)1 controls the sequential progression from the pEMT phase to the redifferentiation phase. Loss of STAT1 prevents redifferentiation. Constitutively active STAT1 allows redifferentiation to occur even when cells are otherwise prevented from progressing beyond the pEMT phase by exogenous activation of Raf. Moreover, tyrosine phosphorylation defective STAT1 partially restored cord formation in such cells, suggesting that STAT1 functions in part as nonnuclear protein mediating signal transduction in this process. Constitutively active or inactive forms of STAT1 did not promote lumen maturation, suggesting this requires a distinct signal.

scholarly journals Galectin-8 induces partial epithelial–mesenchymal transition with invasive tumorigenic capabilities involving a FAK/EGFR/proteasome pathway in Madin–Darby canine kidney cells

2018 ◽  
Vol 29 (5) ◽  
pp. 557-574 ◽  
Author(s):  
Claudia Oyanadel ◽  
Christopher Holmes ◽  
Evelyn Pardo ◽  
Claudio Retamal ◽  
Ronan Shaughnessy ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Plasminogen Activator ◽  
Epithelial Mesenchymal Transition ◽  
Mdck Cells ◽  
Urokinase Plasminogen Activator ◽  
Madin Darby Canine Kidney ◽  
Mesenchymal Transition ◽  
Β1 Integrins ◽  
Darby Canine Kidney ◽  
Canine Kidney

Epithelial cells can acquire invasive and tumorigenic capabilities through epithelial–mesenchymal-transition (EMT). The glycan-binding protein galectin-8 (Gal-8) activates selective β1-integrins involved in EMT and is overexpressed by certain carcinomas. Here we show that Gal-8 overexpression or exogenous addition promotes proliferation, migration, and invasion in nontumoral Madin–Darby canine kidney (MDCK) cells, involving focal-adhesion kinase (FAK)-mediated transactivation of the epidermal growth factor receptor (EGFR), likely triggered by α5β1integrin binding. Under subconfluent conditions, Gal-8–overexpressing MDCK cells (MDCK-Gal-8H) display hallmarks of EMT, including decreased E-cadherin and up-regulated expression of vimentin, fibronectin, and Snail, as well as increased β-catenin activity. Changes related to migration/invasion included higher expression of α5β1 integrin, extracellular matrix-degrading MMP13 and urokinase plasminogen activator/urokinase plasminogen activator receptor (uPA/uPAR) protease systems. Gal-8–stimulated FAK/EGFR pathway leads to proteasome overactivity characteristic of cancer cells. Yet MDCK-Gal-8Hcells still develop apical/basolateral polarity reverting EMT markers and proteasome activity under confluence. This is due to the opposite segregation of Gal-8 secretion (apical) and β1-integrins distribution (basolateral). Strikingly, MDCK-Gal-8Hcells acquired tumorigenic potential, as reflected in anchorage-independent growth in soft agar and tumor generation in immunodeficient NSG mice. Therefore, Gal-8 can promote oncogenic-like transformation of epithelial cells through partial and reversible EMT, accompanied by higher proliferation, migration/invasion, and tumorigenic properties.

Betaine and inositol reduce MDCK cell glycerophosphocholine by stimulating its degradation

1996 ◽  
Vol 270 (1) ◽  
pp. C200-C207 ◽  
Author(s):  
E. D. Kwon ◽  
K. Zablocki ◽  
E. M. Peters ◽  
K. Y. Jung ◽  
A. Garcia-Perez ◽  
...  
Keyword(s):  
Mdck Cell ◽  
Mdck Cells ◽  
Phospholipase Activity ◽  
Madin Darby Canine Kidney ◽  
Data Support ◽  
Compatible Osmolytes ◽  
Darby Canine Kidney ◽  
Canine Kidney ◽  
Medullary Cells

The amount of glycerophosphocholine (GPC) in renal medullary cells in vivo and in cultured renal [Madin-Darby canine kidney (MDCK)] cells varies with extracellular NaCl and urea. We previously showed that this is largely due to modulation of GPC degradation catalyzed by GPC:choline phosphodiesterase (GPC: PDE). GPC also varies inversely with the levels of other compatible osmolytes, the accumulation of which is induced by high tonicity. We tested whether GPC:PDE activity and GPC degradation are affected by accumulation of compatible osmolytes other than GPC. We find that MDCK cell GPC content decreases when the cells take up betaine and/or inositol from the medium. The effect is considerably greater for cells in isosmotic or high-NaCl medium than in high-urea medium. This difference is associated with suppression of betaine and inositol accumulation with high urea. We then measured GPC:PDE activity with a novel chemiluminescent assay. Addition of inositol and/or betaine to the medium greatly increases GPC:PDE activity in cells in isosmotic or high-NaCl media, but the increase is much less in high-urea medium. The increases in GPC:PDE activity, associated with the presence of betaine, are accompanied by commensurate increases in absolute rates of endogenous GPC degradation by cells in isosmotic or high-NaCl medium. We found previously that, in MDCK cells incubated for 2 days in high-NaCl medium, the rate of GPC synthesis from phosphatidylcholine is increased, correlated with an increase in phospholipase activity. However, in the present experiments, betaine accumulation has no effect on phospholipase activity under those conditions and, thus, presumably does not affect GPC synthesis. Collectively, these data support the conclusion that betaine and/or inositol reduces GPC by increasing GPC degradation catalyzed by GPC:PDE. This mechanism enables GPC to be reciprocally regulated relative to other compatible osmolytes, thus maintaining an appropriate total osmolyte content.

scholarly journals Penetration of Salmonella through a polarized Madin-Darby canine kidney epithelial cell monolayer.

1988 ◽  
Vol 107 (1) ◽  
pp. 221-230 ◽  
Author(s):  
B B Finlay ◽  
B Gumbiner ◽  
S Falkow
Keyword(s):  
Mdck Cell ◽  
Pathogenic Bacteria ◽  
Mdck Cells ◽  
Cell Monolayer ◽  
Apical Surface ◽  
Madin Darby Canine Kidney ◽  
Intracellular Parasites ◽  
Epithelial Barriers ◽  
Darby Canine Kidney ◽  
Canine Kidney

Many intracellular parasites are capable of penetrating host epithelial barriers. To study this process in more detail we examined the interactions between the pathogenic bacteria Salmonella choleraesuis and polarized epithelial monolayers of Madin-Darby canine kidney (MDCK) cells grown on membrane filters. Association of bacteria with the MDCK cell apical surface was an active event, requiring bacterial RNA and protein synthesis, and was blocked by low temperatures. Salmonella were internalized within a membrane-bound vacuole and exhibited penetration through, but not between MDCK cells. A maximum of 14 Salmonella per MDCK cell crossed the monolayer per hour to the basolateral surface yet the monolayer remained viable and impermeable to Escherichia coli. Apical S. choleraesuis infection resulted in an increase in paracellular permeability but the MDCK intercellular contacts were not significantly disrupted. Basolateral S. choleraesuis infection was inefficient, and only small numbers of S. choleraesuis penetrated to the apical medium.

scholarly journals Characterization of ZO-1, a protein component of the tight junction from mouse liver and Madin-Darby canine kidney cells.

1988 ◽  
Vol 106 (4) ◽  
pp. 1141-1149 ◽  
Author(s):  
J M Anderson ◽  
B R Stevenson ◽  
L A Jesaitis ◽  
D A Goodenough ◽  
M S Mooseker
Keyword(s):  
Tight Junction ◽  
Mdck Cell ◽  
Mouse Liver ◽  
Mdck Cells ◽  
Cell Protein ◽  
Scatchard Analysis ◽  
Madin Darby Canine Kidney ◽  
Mouse Tissues ◽  
Darby Canine Kidney ◽  
Canine Kidney

ZO-1, originally identified by mAb techniques, is the first protein shown to be specifically associated with the tight junction. Here we describe and compare the physical characteristics of ZO-1 from mouse liver and the Madin-Darby canine kidney (MDCK) epithelial cell line. The ZO-1 polypeptide has an apparent size of 225 kD in mouse tissues and 210 kD in canine-derived MDCK cells as determined by SDS-PAGE/immunoblot analysis. ZO-1 from both sources is optimally solubilized from isolated plasma membranes by either 6 M urea or high pH conditions; partial solubilization occurs with 0.3 M KCl. The nonionic detergents, Triton X-100 and octyl-beta-D-glucopyranoside, do not solubilize ZO-1. These solubility properties indicate that ZO-1 is a peripherally associated membrane protein. ZO-1 was purified to electrophoretic homogeneity from [35S]methionine metabolically labeled MDCK cells by a combination of gel filtration and immunoaffinity chromatography. Purified ZO-1 has an s20,w of 5.3 and Stokes radius of 8.6 nm. These values suggest that purified ZO-1 is an asymmetric monomeric molecule. Corresponding values for mouse liver ZO-1, characterized in impure protein extracts, were 6 s20,w and 9 nm. ZO-1 was shown to be a phosphoprotein in MDCK cells metabolically labeled with [32P]orthophosphate; analysis of phosphoamino acids from purified ZO-1 revealed only phosphoserine. ZO-1 epitope number was determined by Scatchard analysis of competitive and saturable binding of two different 125I-mAbs to SDS-solubilized proteins from liver and MDCK cells immobilized on nitrocellulose. Saturation binding occurs at 26 ng mAb/mg liver and 63 ng/mg of MDCK cell protein. This is equivalent to 30,000 ZO-1 molecules per MDCK cell assuming a single epitope/ZO-1 molecule.

Sign in / Sign up

Export Citation Format

Share Document