Effect of Wnt/β-catenin and NF-κB signaling pathways on mucus secretion with hypertonicity in 16HBE cells

Abstract

This study aimed at identifying the molecular mechanisms and effects of hypertonicity on mucin5AC (MUC5AC) expression in airway epithelial cells for which immortalized human bronchial epithelial (16HBE) cells were cultured in 600 mOsm/L hypertonic medium for different times in vitro. Proteins of MUC5AC and β-catenin, Cyclin D1, NF-κB p65 were detected by enzyme linked immunosorbent assay (ELISA), reverse transcription (RT)-PCR and western blotting assays. After transfection of β-catenin siRNA in 16HBE cells, the levels of β-catenin, Cyclin D1, NF-κB p65 and MUC5AC protein were detected. Results showed that the levels of mRNAs and proteins of target genes increased significantly after the exposure to hypertonic conditions and the expression content increased in a time-dependent manner. Furthermore, transfection with β-catenin siRNA attenuated the expression of these genes. These results suggested that the Wnt/β-catenin and NF-κB signaling pathways played essential roles in inducing the MUC5AC hypersecretion in 16HBE cells in response to hypertonicity.

Keywords

• MUC5AC
• hypertonicity
• Wnt/β-catenin
• NF-κB p65