Notch signaling regulates col1α1 and col1α2 expression in airway fibroblasts.

Abstract

Subepithelial fibrosis is one of the common pathological features of asthmatic airway remodeling. During subepithelial fibrosis, type I collagen becomes the most abundant extracellular protein component. Studies have shown that Notch signaling participates in the progression of fibrosis; however, whether Notch signaling is involved in regulating type I collagen expression in airway fibroblasts remains unclear. The aim of the present study was to examine whether Notch signaling can regulate type I collagen expression in airway fibroblasts and to explore the underlying molecular mechanisms. Here, the expression of Notch signaling components was examined in mouse L929 cells and human MRC-5 cells. After upregulating or downregulating Notch signaling in these cell lines, col1α1 and col1α2 expression was examined. Using gene reporter assays, site-directed mutagenesis, and ChIP assays, the role of Hes1 binding sites in both the mouse and human COL1A1 and COL1A2 promoters was investigated. This study revealed that Notch signaling-related molecules (including Notch1, Hes1, and others) are expressed in L929 and MRC-5 cells and that Notch signaling regulates the expression of col1α1 and col1α2 in both cell lines. Additionally, over-expression of the Notch intracellular domain resulted in activation of the COL1A1 and COL1A2 promoters, and site-directed mutagenesis reporter assays revealed that Hes1 proteins might augment both mouse and human COL1A1 and COL1A2 promoter activity. Furthermore, ChIP assays confirmed that Hes1 binds to the COL1A1 and COL1A2 promoters in both L929 and MRC-5 cells. Therefore, it is reasonable to assume that Notch signaling can directly upregulate COL1A1 and COL1A2 promoter activity through a Hes1-dependent mechanism, which could serve as a possible target for pharmacotherapy of airway subepithelial fibrosis.

• Authors: Mei Hu, Hai-Feng Ou-Yang, Chang-Gui Wu, Shuo-Yao Qu, Xin-Ting Xu, Ping Wang
• Journal: Experimental biology and medicine (Maywood, N.J.) (Exp Biol Med (Maywood)) Vol. 239 Issue 12 Pg. 1589-96 (Dec 2014) ISSN: 1535-3699 [Electronic] England
• PMID: 25107895 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 by the Society for Experimental Biology and Medicine.

Chemical References

• Basic Helix-Loop-Helix Transcription Factors
• Collagen Type I
• Hes1 protein, mouse
• Homeodomain Proteins
• Receptors, Notch
• Transcription Factor HES-1
• HES1 protein, human

Topics

• Animals
• Basic Helix-Loop-Helix Transcription Factors (metabolism)
• Binding Sites
• Cell Line
• Chromatin Immunoprecipitation
• Collagen Type I (biosynthesis)
• Fibroblasts (physiology)
• Gene Expression Regulation
• Genes, Reporter
• Homeodomain Proteins (metabolism)
• Humans
• Mice
• Mutagenesis, Site-Directed
• Promoter Regions, Genetic
• Protein Binding
• Receptors, Notch (metabolism)
• Signal Transduction
• Transcription Factor HES-1