TGF-β1 Induces Human Bronchial Epithelial Cell-to-Mesenchymal Transition in Vitro
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The subepithelial fibrosis component of airway remodeling in asthma is mediated through induction of transforming growth factor-β1 (TGF-β1) expression with consequent activation of myofibroblasts to produce extracellular matrix proteins. The number of myofibroblasts is increased in the asthmatic airway and is significantly correlated with the thickness of lamina reticularis. However, much is still unknown regarding the origin of bronchial myofibroblasts. Emerging evidence suggests that myofibroblasts can derive from epithelial cells by an epithelial-to-mesenchymal transition (EMT). In this study we investigated whether TGF-β1 could induce bronchial epithelial EMT in the human bronchial epithelial cell. Cultured human bronchial epithelial cells, 16HBE-14o, were stimulated with 10 ng/ml TGF-β1. Morphologic changes were observed and stress fiber by actin reorganization was detected by indirect immunostaining. The expression of α-SMA (α-smooth muscle actin) and the epithelial cell marker E-cadherin were detected in those 16HBE-14o cells after TGF-β1 stimulation for 72 h, using immunostaining and RT-PCR. The contents of collagen I were determined by radioimmunoassay, and the levels of endogenous TGF-β1 were measured with ELISA. Human bronchial epithelial cells stimulated with TGF-β1 were converted from a “cobblestone” epithelial structure into an elongated fibroblast-like shape. Incubation of human bronchial epithelial cells with TGF-β1 induced de novo expression of α-SMA, increased formation of stress fiber by F-actin reorganization, and loss of epithelial marker E-cadherin. Moreover, a significant increase in the levels of collagen I and endogenous TGF-β1 released from bronchial epithelial cells stimulated with TGF-β1 were observed. These results suggested that human bronchial epithelial cells, under stimulation of TGF-β1, underwent transdifferentiation into myofibroblasts.
KeywordsBronchial epithelial cell Transforming growth factor-β1 Epithelial-to-mesenchymal transition
This work was supported by Guangzhou Medical and Health Science Research Foundation (No. 2007-ZDi-06, No. 2007-YB-040) and Guangdong Medical Science Research Foundation (No. A2008537).
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