Chloe Leduc1, Lauren Sobilo2, Hechmi Toumi3, Philippe Mondon4, Eric Lespessailles3, Fédéric Ossant5, Robin Kurfurst2, Chantal Pichon6. 1. Centre de Biophysique Moléculaire, CNRS UPR 4301, University of Orléans, Orléans, France. 2. LVMH Recherche, 185 avenue de Verdun, Saint Jean de Braye, France. 3. I3MTO EA 4708, University of Orléans, Orléans, France. 4. Sederma, 29 Rue du Chemin Vert, 78610 Le Perray en Yvelines, France. 5. Inserm U930, Université de Tours, UFR Médecine, 2 Boulevard Tonnellé, 37044 Tours, France. 6. Centre de Biophysique Moléculaire, CNRS UPR 4301, University of Orléans, Orléans, France. Electronic address: pichon@cnrs-orleans.fr.
Abstract
BACKGROUND: Transforming growth factor beta inducible early gene-1 (TIEG-1), a member of the Krüppel-like factor, was identified as a primary response gene for TGF-β. The role of TIEG-1 in skin repair has been mainly addressed in vivo on TIEG-1 null mice model and the mechanism remains unexplored. METHODS: We investigated the modulation of TIEG-1 expression in normal human skin fibroblasts by either down-expressing or overexpressing the gene. We evaluated reactive oxygen species production and the cell viability of treated cells. The effect of TIEG-1 overexpression was monitored by wound healing assay and immunofluorescence staining of actin fibers organization and alpha-smooth muscle actin (α-SMA). Western blots were carried out to identify the level of expression or phosphorylation of key proteins such as cofilin, Rho GTPases, and p38 mitogen-activated protein kinase (p38 MAPK). RESULTS: TIEG-1 down-regulation had a deleterious effect on the cell viability. It was significantly reduced (65±5%) and exposure to ultraviolet further increased this effect (47±3%). By contrast, cells overexpressing TIEG-1 had a reduced reactive oxygen species production (75%) compared to control and mock-transfected cells. This overexpression also resulted in formation of actin stress fibers and increased α-SMA expression and an enhanced wound healing feature. RhoB GTPase was upregulated and phosphorylation of cofilin and p38 MAPK was observed. CONCLUSION: TIEG-1 overexpression in normal human skin fibroblasts results in improved resistance to oxidative stress, myofibroblast-like conversion that involved RhoB signaling pathway with cofilin and p38 MAPK proteins activation. GENERAL SIGNIFICANCE: This study enlightens the role of TIEG-1 role in skin biology.
BACKGROUND: Transforming growth factor beta inducible early gene-1 (TIEG-1), a member of the Krüppel-like factor, was identified as a primary response gene for TGF-β. The role of TIEG-1 in skin repair has been mainly addressed in vivo on TIEG-1 null mice model and the mechanism remains unexplored. METHODS: We investigated the modulation of TIEG-1 expression in normal human skin fibroblasts by either down-expressing or overexpressing the gene. We evaluated reactive oxygen species production and the cell viability of treated cells. The effect of TIEG-1 overexpression was monitored by wound healing assay and immunofluorescence staining of actin fibers organization and alpha-smooth muscle actin (α-SMA). Western blots were carried out to identify the level of expression or phosphorylation of key proteins such as cofilin, Rho GTPases, and p38 mitogen-activated protein kinase (p38 MAPK). RESULTS:TIEG-1 down-regulation had a deleterious effect on the cell viability. It was significantly reduced (65±5%) and exposure to ultraviolet further increased this effect (47±3%). By contrast, cells overexpressing TIEG-1 had a reduced reactive oxygen species production (75%) compared to control and mock-transfected cells. This overexpression also resulted in formation of actin stress fibers and increased α-SMA expression and an enhanced wound healing feature. RhoB GTPase was upregulated and phosphorylation of cofilin and p38 MAPK was observed. CONCLUSION:TIEG-1 overexpression in normal human skin fibroblasts results in improved resistance to oxidative stress, myofibroblast-like conversion that involved RhoB signaling pathway with cofilin and p38 MAPK proteins activation. GENERAL SIGNIFICANCE: This study enlightens the role of TIEG-1 role in skin biology.
Authors: Sarah K Abey; Yuana Yuana; Paule V Joseph; Natnael D Kenea; Nicolaas H Fourie; LeeAnne B Sherwin; Gregory E Gonye; Paul A Smyser; Erin S Stempinski; Christina M Boulineaux; Kristen R Weaver; Christopher K E Bleck; Wendy A Henderson Journal: BBA Clin Date: 2016-12-20
Authors: Yasin Shaifta; Charles E MacKay; Nneka Irechukwu; Katie A O'Brien; David B Wright; Jeremy P T Ward; Greg A Knock Journal: J Physiol Date: 2017-11-23 Impact factor: 5.182