scholarly journals Explanting Is anEx VivoModel of Renal Epithelial-Mesenchymal Transition

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Catherine E. Winbanks ◽  
Ian A. Darby ◽  
Kristen J. Kelynack ◽  
Dodie Pouniotis ◽  
Gavin J. Becker ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
De Novo ◽  
Epithelial Mesenchymal Transition ◽  
Ex Vivo ◽  
Rat Kidney ◽  
Smooth Muscle Actin ◽  
Muscle Actin ◽  
Alpha Smooth Muscle Actin ◽  
Mesenchymal Transition ◽  
Renal Tubulointerstitial Fibrosis

Recognised by theirde novoexpression of alpha-smooth muscle actin (SMA), recruitment of myofibroblasts is key to the pathogenesis of fibrosis in chronic kidney disease. Increasingly, we realise that epithelial-mesenchymal transition (EMT) may be an important source of these cells. In this study we describe a novel model of renal EMT. Rat kidney explants were finely diced on gelatin-coated Petri dishes and cultured in serum-supplemented media. Morphology and immunocytochemistry were used to identify mesenchymal (vimentin+, α-smooth muscle actin (SMA)+, desmin+), epithelial (cytokeratin+), and endothelial (RECA+) cells at various time points. Cell outgrowths were all epithelial in origin (cytokeratin+) at day 3. By day 10, 50 ± 12% (mean ± SE) of cytokeratin+ cells double-labelled for SMA, indicating EMT. Lectin staining established a proximal tubule origin. By day 17, cultures consisted only of myofibroblasts (SMA+/cytokeratin−). Explanting is a reproducibleex vivomodel of EMT. The ability to modify this change in phenotype provides a useful tool to study the regulation and mechanisms of renal tubulointerstitial fibrosis.

Differential regulation of E-cadherin and α-smooth muscle actin by BMP 7 in human renal proximal tubule epithelial cells and its implication in renal fibrosis

2009 ◽  
Vol 297 (5) ◽  
pp. F1238-F1248 ◽  
Author(s):  
Mangalakumar Veerasamy ◽  
Tri Q. Nguyen ◽  
Reza Motazed ◽  
Alexander L. Pearson ◽  
Roel Goldschmeding ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Proximal Tubule ◽  
Crucial Role ◽  
De Novo ◽  
Epithelial Mesenchymal Transition ◽  
Smooth Muscle Actin ◽  
Cell Phenotype ◽  
Muscle Actin ◽  
Mesenchymal Transition ◽  
E Cadherin

Chronic kidney diseases are characterized by progressive tubulointerstitial fibrosis, and TGFβ1 plays a crucial role in its development. Bone morphogenic protein 7 (BMP 7), another member of the TGF superfamily, antagonized the profibrotic effects of TGFβ1, including epithelial mesenchymal transition and E-cadherin loss, in the previous studies from animal models. We investigated the effect of BMP 7 on TGFβ1-mediated E-cadherin loss in two different transformed human adult proximal tubule epithelia. We found that BMP 7 not only failed to prevent TGFβ1-mediated E-cadherin loss but itself downregulated E-cadherin levels and that it had an additive effect with TGFβ1 in inducing E-cadherin loss. The downregulation of E-cadherin by BMP 7 was mediated through the Smad1/5 pathway. BMP 7-mediated E-cadherin loss was not followed by de novo α-smooth muscle actin (α-SMA) expression (a marker of myofibroblastic phenotype), which was due to the concurrent induction of Inhibitor of DNA binding 1 (Id1, a basic helix loop helix class transcriptional regulator) through a non-Smad pathway. Concurrent treatment of BMP 7 and TGFβ1 prevented TGFβ1-mediated α-SMA induction. In summary, our results suggest that E-cadherin loss, the key feature of epithelial mesenchymal transition, will not necessarily be followed by total phenotype change; rather, cells may undergo some loss of phenotypic marker in a ligand-dependent manner and participate in reparative processes. The inhibition of de novo expression of α-SMA could explain the antifibrotic effect of BMP 7. Id1 might play a crucial role in maintaining proximal tubule epithelial cell phenotype and its signaling regulation could be a potential therapeutic target.

255 GAMMA SMOOTH MUSCLE ACTIN: AN EPITHELIAL MESENCHYMAL TRANSITION MARKER IN HEPATOCELLULAR CARCINOMA

2012 ◽  
Vol 56 ◽  
pp. S107
Author(s):  
N. Benzoubir ◽  
A. Dos Santos ◽  
C. Guillaume ◽  
D. Samuel ◽  
C. Brechot ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Smooth Muscle ◽  
Epithelial Mesenchymal Transition ◽  
Smooth Muscle Actin ◽  
Muscle Actin ◽  
Mesenchymal Transition

scholarly journals Morphological Changes in the Cells of the Ductal Adenocarcinoma of the Pancreas at Epithelial-Mesenchymal Transition

2019 ◽  
Vol 8 (3) ◽  
pp. 59-65
Author(s):  
T. N. Sotnikova ◽  
G. R. Setdikova ◽  
O. V. Paklina ◽  
V. P. Shubin ◽  
M. V. Mnikhovich ◽  
...  
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Epithelial Mesenchymal Transition ◽  
Smooth Muscle Actin ◽  
Molecular Genetic ◽  
Ductal Adenocarcinoma ◽  
Marker Genes ◽  
Muscle Actin ◽  
Mesenchymal Transition ◽  
Adenocarcinoma Of The Pancreas

The aim of the research was to study the expression of marker genes for the epithelial-mesenchymal transition in the ductal adenocarcinoma of the pancreas.Material and methods. Surgical material from 44 patients with ductal adenocarcinoma of the pancreas was subjected to morphological analysis with molecular genetic research. Total RNA was detected in the detected sections of the anaplastic component using the RNeasy Mini Kit (Qiagen, Germany). 5 marker genes were used for molecular genetic studies of the epithelial-mesenchymal transition (EMT): ZEB1, ZEB2, CDH1, VIM, SNAIL1 (SLUG). Gene expression was measured in triplicate using an EvaGreen intercalating dye. On serial paraffin sections using tissue microarrays technology, immunohistochemical detection of p63, smooth muscle actin, total cytokeratin, cytokeratin 7, vimentin, E-cadherin (Ventana) was performed.Results. As a result of the study, a positive reaction with mesenchymal markers (vimentin, p63, smooth muscle actin) was detected in the cells of the anaplastic component, in contrast to the glandular component. In a molecular study of the anaplastic component, changes in gene expression were characterized as EMT-positive.Conclusion. The heterogeneity of ductal cancer is manifested in the appearance of an anaplastic (sarcomalike) component, in which the ability of epithelial tumor cells to acquire the property of mesenchymal cells that do not require stroma and have aggressive malignant potential that affects the survival of patients is traced.

Central role for Rho in TGF-β1-induced α-smooth muscle actin expression during epithelial-mesenchymal transition

2003 ◽  
Vol 284 (5) ◽  
pp. F911-F924 ◽  
Author(s):  
András Masszi ◽  
Caterina Di Ciano ◽  
Gábor Sirokmány ◽  
William T. Arthur ◽  
Ori D. Rotstein ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Transforming Growth Factor ◽  
Epithelial Mesenchymal Transition ◽  
Smooth Muscle Actin ◽  
Stress Fiber ◽  
Small Gtpase ◽  
Cell Contact ◽  
Muscle Actin ◽  
Mesenchymal Transition ◽  
Fiber Assembly

New research suggests that, during tubulointerstitial fibrosis, α-smooth muscle actin (SMA)-expressing mesenchymal cells might derive from the tubular epithelium via epithelial-mesenchymal transition (EMT). Although transforming growth factor-β1(TGF-β1) plays a key role in EMT, the underlying cellular mechanisms are not well understood. Here we characterized TGF-β1-induced EMT in LLC-PK1 cells and examined the role of the small GTPase Rho and its effector, Rho kinase, (ROK) in the ensuing cytoskeletal remodeling and SMA expression. TGF-β1 treatment caused delocalization and downregulation of cell contact proteins (ZO-1, E-cadherin, β-catenin), cytoskeleton reorganization (stress fiber assembly, myosin light chain phosphorylation), and robust SMA synthesis. TGF-β1induced a biphasic Rho activation. Stress fiber assembly was prevented by the Rho-inhibiting C3 transferase and by dominant negative (DN) ROK. The SMA promoter was activated strongly by constitutively active Rho but not ROK. Accordingly, TGF-β1-induced SMA promoter activation was potently abrogated by two Rho-inhibiting constructs, C3 transferase and p190RhoGAP, but not by DN-ROK. Truncation analysis showed that the first CC(A/T)richGG (CArG B) serum response factor-binding cis element is essential for the Rho responsiveness of the SMA promoter. Thus Rho plays a dual role in TGF-β1-induced EMT of renal epithelial cells. It is indispensable both for cytoskeleton remodeling and for the activation of the SMA promoter. The cytoskeletal effects are mediated via the Rho/ROK pathway, whereas the transcriptional effects are partially ROK independent.

scholarly journals Inhibition of TGF-β2-induced migration and epithelial-mesenchymal transition in ARPE-19 by sulforaphane

2021 ◽  
Vol 14 (7) ◽  
pp. 973-980
Author(s):  
Yan-Bing Huang ◽  
◽  
Hui Zheng ◽  
Xiu-Xia Yang ◽  
Cheng-Cheng Yang ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Transforming Growth Factor ◽  
Epithelial Mesenchymal Transition ◽  
Smooth Muscle Actin ◽  
Fiber Formation ◽  
Stress Fibers ◽  
Actin Stress Fiber ◽  
Muscle Actin ◽  
Mesenchymal Transition ◽  
Actin Stress Fibers

AIM: To investigate the effects of sulforaphane (SFN) on transforming growth factor (TGF)-β2 stimulated migration and epithelial-mesenchymal transition (EMT) in ARPE-19 cells. METHODS: ARPE-19 cells were cultured in the presence or absence of SFN or TGF-β2. SFN toxicity was assessed by performing a lactate dehydrogenase assay (LDH) and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assays, and cell migration was evaluated by Transwell migration assay. Actin stress fiber formation in ARPE-19 cells was determined using immunofluorescence analysis. Immunoblotting analysis was used to determine fibronectin and α-smooth muscle actin expressions along with the degree of Smad and Akt phosphorylation. RESULTS: SFN inhibited ARPE-19 migration. Additionally, SFN attenuated TGF-β2-induced appearance of actin stress fibers as well as fibronectin and α-smooth muscle actin expressions in these cells. SFN also hindered the TGF-β2-stimulated phosphorylation of Smad2, Smad3, and Akt. SFN showed no cytotoxicity towards ARPE-19 cells. CONCLUSION: SFN inhibits TGF-β2-stimulated migration and EMT in ARPE-19 cells, probably by preventing the establishment of actin stress fibers and Akt and Smad2/3 signaling.

scholarly journals Gamma-Smooth Muscle Actin Expression Is Associated with Epithelial-Mesenchymal Transition and Stem-Like Properties in Hepatocellular Carcinoma

PLoS ONE ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. e0130559 ◽  
Author(s):  
Nassima Benzoubir ◽  
Charlotte Mussini ◽  
Charlène Lejamtel ◽  
Alexandre Dos Santos ◽  
Claire Guillaume ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Smooth Muscle ◽  
Epithelial Mesenchymal Transition ◽  
Smooth Muscle Actin ◽  
Muscle Actin ◽  
Mesenchymal Transition ◽  
Actin Expression
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