TGF-β1 targets the GSK-3β/β-catenin pathway via ERK activation in the transition of human lung fibroblasts into myofibroblasts

Transforming growth factor-β1 (TGF-β1) is known to induce the transition of human lung fibroblasts to myofibroblasts, a primary event in the pathogenesis of idiopathic pulmonary fibrosis. The molecular pathways involved in myofibroblast transformation are only partially identified.We found that a 24-h treatment with TGF-β1 (10 ng/ml) induced α-smooth actin (SMA) expression and collagen production in human lung fibroblasts. These effects were abrogated by PD98059, a specific inhibitor of the mitogen-activated protein kinase (MAPK) pathway. TGF-β1 treatment activated the MAPK pathway, as shown by an increased phosphorylation of extracellular-regulated kinases (ERK)1/2 after 30 min of exposure. TGF-β1 also increased the expression of the Ser-9-phosphorylated inactive form of glycogen synthase kinase-3β (GSK-3β), an effect that was largely attenuated by PD98059. A nuclear translocation of β-catenin in human lung fibroblasts was observed 2 h after TGF-β1 addition both by confocal microscopy and nuclear protein analysis. At this time, TGF-β1 also increased the total levels of β-catenin, an effect that was prevented by PD98059. Similarly to TGF-β1, the GSK-3β inhibitor lithium chloride (10 mM), increased the total levels of β-catenin and promoted α-SMA expression and collagen production.This study demonstrates that TGF-β1 induces α-SMA expression and collagen production in human lung fibroblasts via ERK1/2 activation, GSK-3β inhibition and nuclear β-catenin translocation. The evidence that the silencing of β-catenin by siRNAs was able to prevent the induction of α-SMA expression in TGF-β1-treated fibroblasts further supports the hypothesis of a contribution of the GSK-3β/β-catenin pathway in the pathogenesis of idiopathic pulmonary fibrosis.