Human Neutrophil Elastase Induces MUC5AC Overexpression in Chronic Rhinosinusitis Through miR-146a.

Abstract

Background The pathogenesis of chronic rhinosinusitis (CRS) is not yet clear. microRNAs are widely involved in a number of physiological and pathological processes, of which microRNA-146a (miR-146a) plays an important role in innate immunity, inflammatory response, and other pathophysiological processes. Mucins (MUCs) are important components of secreted mucus, of which MUC5AC is the major MUC secreted in the normal airway. Objective This study was performed to examine human neutrophil elastase (HNE)-induced MUC5AC overexpression in CRS via miR-146a. Methods miR-146a, HNE, epidermal growth factor receptor (EGFR), and MUC5AC expression in the sinonasal mucosa were determined using quantitative real-time polymerase chain reaction (qRT-PCR). EGFR, phosphorylated EGFR (pEGFR), and MUC5AC expression were determined in primary cultures of human nasal epithelial cells (HNECs). We examined the expression of miR-146a, MUC5AC, EGFR, and pEGFR by transfecting HNECs with miR-146a mimics and negative control (NC). Moreover, dual-luciferase reporter gene assays were used to validate EGFR as an hsa-miR-146a target gene. Results miR-146a was significantly downregulated, and HNE, EGFR, and MUC5AC were upregulated in CRS patients both with and without nasal polyps. In the in vitro cell experiment, MUC5AC was significantly downregulated after use of an EGFR-specific inhibitor (AG1478). Upon addition of miR-146a inhibitor, miR-146a was downregulated, while MUC5AC was upregulated. MUC5AC was suppressed in normal primary HNECs by miR-146a mimic and pEGFR was downregulated. The results of dual-luciferase reporter assays showed that the luciferase activities were markedly inhibited in the pGL3-EGFR-3′ UTR+miR-146a mimic group compared with the pGL3+ miR-146a mimic group, suggesting that EGFR is a target gene for miR-146a. Conclusion In HNE-induced CRS, miR-146a downregulates the expression of MUC5AC by inhibiting the activation of EGFR, and EGFR is a target gene of miR-146a.