DNMTs Are Involved in TGF-β1-Induced Epithelial-Mesenchymal Transitions in Airway Epithelial Cells

Chronic rhinosinusitis (CRS) pathogenesis is closely related to tissue remodeling, including epithelial-mesenchymal transition (EMT). Epigenetic mechanisms play key roles in EMT. DNA methylation, mediated by DNA methyltransferases (DNMTs), is an epigenetic marker that is critical to EMT. The goal of this study was to determine whether DNMTs were involved in TGF-beta 1-induced EMT and elucidate the underlying mechanisms in nasal epithelial cells and air-liquid interface cultures. Global DNA methylation and DNMT activity were quantified. DNMT expression was measured using real-time PCR (qRT-PCR) in human CRS tissues. mRNA and protein levels of DNMTs, E-cadherin, vimentin, alpha-SMA, and fibronectin were determined using RT-PCR and Western blotting, respectively. DNMT1, DNMT3A, and DNMT3B gene expression were knocked down using siRNA transfection. MAPK phosphorylation and EMT-related transcription factor levels were determined using Western blotting. Signaling pathways were analyzed using specific inhibitors of MAPK. We demonstrated these data in primary nasal epithelial cells and air-liquid interface cultures. Global DNA methylation, DNMT activity, and DNMT expression increased in CRS tissues. DNMT expression was positively correlated with Lund-McKay CT scores. TGF-beta 1 dose-dependently induced DNMT expression. Further, 5-Aza inhibited TGF-beta 1-induced DNMT, Snail, and Slug expression related to EMT, as well as p38 and JNK phosphorylation in A549 cells and TGF-beta 1-induced DNMT expression and EMT in primary nasal epithelial cells and air-liquid interface cultures. TGF-beta 1-induced DNMT expression leads to DNA methylation and EMT via p38, JNK, Snail, and Slug signaling pathways. Inhibition of DNMT suppressed the EMT process and therefore is potentially a CRS therapeutic strategy.

Automated summary

The pathogenesis of chronic rhinosinusitis is closely related to tissue remodeling, particularly epithelial-mesenchymal transition. Epigenetic mechanisms, such as DNA methylation, mediated by DNA methyltransferases, play key roles in this process. This study aimed to investigate the involvement of DNA methyltransferases in TGF-beta 1-induced epithelial-mesenchymal transition and explore the underlying mechanisms. The results showed that DNA methyltransferases were upregulated in chronic rhinosinusitis tissues and their expression was positively correlated with disease severity. Further experiments revealed that TGF-beta 1 induced the expression of DNA methyltransferases, which led to DNA methylation and epithelial-mesenchymal transition via the p38, JNK, Snail, and Slug signaling pathways. Inhibition of DNA methyltransferases suppressed the epithelial-mesenchymal transition process, suggesting its potential as a therapeutic strategy for chronic rhinosinusitis.