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

机译:Wnt /β-catenin和NF-κB信号通路对16HBE细胞高渗性黏液分泌的影响

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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.
机译:这项研究旨在确定高渗性对气道上皮细胞黏蛋白5AC(MUC5AC)表达的分子机制和影响,在600 mOsm / L高渗培养基中体外培养不同时间的永生化人支气管上皮细胞(16HBE)。通过酶联免疫吸附法(ELISA),逆转录(RT)-PCR和western blotting法检测MUC5AC和β-catenin,Cyclin D1,NF-κBp65蛋白。在16HBE细胞中转染β-cateninsiRNA后,检测了β-catenin,Cyclin D1,NF-κBp65和MUC5AC蛋白的水平。结果表明,高渗条件下,靶基因的mRNA和蛋白质水平显着增加,表达量呈时间依赖性。此外,用β-cateninsiRNA转染可减弱这些基因的表达。这些结果表明,Wnt /β-catenin和NF-κB信号通路在诱导高渗性的16HBE细胞中MUC5AC过度分泌中起重要作用。

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