SMAD inhibition attenuates epithelial to mesenchymal transition by primary keratinocytes in vitro
Epithelial to mesenchymal transition (EMT) is a process whereby epithelial cells undergo transition to a mesenchymal phenotype and contribute directly to fibrotic disease. Recent studies support a role for EMT in cutaneous fibrotic diseases including scleroderma and hypertrophic scarring, although t...
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Published in | Experimental dermatology Vol. 23; no. 7; pp. 497 - 503 |
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Main Authors | , , , , , , , , , , |
Format | Journal Article |
Language | English |
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Blackwell Publishing Ltd
01.07.2014
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Abstract | Epithelial to mesenchymal transition (EMT) is a process whereby epithelial cells undergo transition to a mesenchymal phenotype and contribute directly to fibrotic disease. Recent studies support a role for EMT in cutaneous fibrotic diseases including scleroderma and hypertrophic scarring, although there is limited data on the cytokines and signalling mechanisms regulating cutaneous EMT. We investigated the ability of TGF‐β and TNF‐α, both overexpressed in cutaneous scleroderma and central mediators of EMT in other epithelial cell types, to induce EMT in primary keratinocytes and studied the signalling mechanisms regulating this process. TGF‐β induced EMT in normal human epidermal keratinocytes (NHEK cells), and this process was enhanced by TNF‐α. EMT was characterised by changes in morphology, proteome (down‐regulation of E‐cadherin and Zo‐1 and up‐regulation of vimentin and fibronectin), MMP secretion and COL1α1 mRNA expression. TGF‐β and TNF‐α in combination activated SMAD and p38 signalling in NHEK cells. P38 inhibition with SB203580 partially attenuated EMT, whereas SMAD inhibition using SB431542 significantly inhibited EMT and also reversed established EMT. These data highlight the retained plasticity of adult keratinocytes and support further studies of EMT in clinically relevant in vivo models of cutaneous fibrosis and investigation of SMAD inhibition as a potential therapeutic intervention. |
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AbstractList | Epithelial to mesenchymal transition (EMT) is a process whereby epithelial cells undergo transition to a mesenchymal phenotype and contribute directly to fibrotic disease. Recent studies support a role for EMT in cutaneous fibrotic diseases including scleroderma and hypertrophic scarring, although there is limited data on the cytokines and signalling mechanisms regulating cutaneous EMT. We investigated the ability of TGF-β and TNF-α, both overexpressed in cutaneous scleroderma and central mediators of EMT in other epithelial cell types, to induce EMT in primary keratinocytes and studied the signalling mechanisms regulating this process. TGF-β induced EMT in normal human epidermal keratinocytes (NHEK cells), and this process was enhanced by TNF-α. EMT was characterised by changes in morphology, proteome (down-regulation of E-cadherin and Zo-1 and up-regulation of vimentin and fibronectin), MMP secretion and COL1α1 mRNA expression. TGF-β and TNF-α in combination activated SMAD and p38 signalling in NHEK cells. P38 inhibition with SB203580 partially attenuated EMT, whereas SMAD inhibition using SB431542 significantly inhibited EMT and also reversed established EMT. These data highlight the retained plasticity of adult keratinocytes and support further studies of EMT in clinically relevant in vivo models of cutaneous fibrosis and investigation of SMAD inhibition as a potential therapeutic intervention. Epithelial to mesenchymal transition ( EMT ) is a process whereby epithelial cells undergo transition to a mesenchymal phenotype and contribute directly to fibrotic disease. Recent studies support a role for EMT in cutaneous fibrotic diseases including scleroderma and hypertrophic scarring, although there is limited data on the cytokines and signalling mechanisms regulating cutaneous EMT . We investigated the ability of TGF ‐ β and TNF ‐ α , both overexpressed in cutaneous scleroderma and central mediators of EMT in other epithelial cell types, to induce EMT in primary keratinocytes and studied the signalling mechanisms regulating this process. TGF‐ β induced EMT in normal human epidermal keratinocytes ( NHEK cells), and this process was enhanced by TNF ‐ α . EMT was characterised by changes in morphology, proteome (down‐regulation of E‐cadherin and Zo‐1 and up‐regulation of vimentin and fibronectin), MMP secretion and COL 1 α 1 mRNA expression. TGF‐ β and TNF‐ α in combination activated SMAD and p38 signalling in NHEK cells. P38 inhibition with SB203580 partially attenuated EMT , whereas SMAD inhibition using SB431542 significantly inhibited EMT and also reversed established EMT . These data highlight the retained plasticity of adult keratinocytes and support further studies of EMT in clinically relevant in vivo models of cutaneous fibrosis and investigation of SMAD inhibition as a potential therapeutic intervention. |
Author | Spence, Shaun O'Kane, Cecilia M. Denton, Christopher P. O'Kane, Donal Damkat-Thomas, Lindsay McAuley, Daniel F. Smyth, Anita E. Jackson, Megan V. Tolland, Julia P. Kissenpfennig, Adrien Stuart Elborn, J. |
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BackLink | https://www.ncbi.nlm.nih.gov/pubmed/24848428$$D View this record in MEDLINE/PubMed |
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Keywords | epithelial to mesenchymal transition keratinocyte transforming growth factor-β tumor necrosis factor-α |
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Snippet | Epithelial to mesenchymal transition (EMT) is a process whereby epithelial cells undergo transition to a mesenchymal phenotype and contribute directly to... Epithelial to mesenchymal transition ( EMT ) is a process whereby epithelial cells undergo transition to a mesenchymal phenotype and contribute directly to... |
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SubjectTerms | Benzamides - chemistry Collagen - metabolism Cytokines - metabolism Dioxoles - chemistry Down-Regulation Epidermis - cytology epithelial to mesenchymal transition Epithelial-Mesenchymal Transition Fibronectins - metabolism Fibrosis - metabolism Humans keratinocyte Keratinocytes - cytology Matrix Metalloproteinases - metabolism p38 Mitogen-Activated Protein Kinases - metabolism Recombinant Proteins - metabolism Signal Transduction Smad Proteins - antagonists & inhibitors Smad Proteins - metabolism Transforming Growth Factor beta - metabolism transforming growth factor-β Tumor Necrosis Factor-alpha - metabolism tumor necrosis factor-α Vimentin - metabolism |
Title | SMAD inhibition attenuates epithelial to mesenchymal transition by primary keratinocytes in vitro |
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