1. Academic Validation
  2. Mucin 1 downregulation impairs the anti-necroptotic effects of glucocorticoids in human bronchial epithelial cells

Mucin 1 downregulation impairs the anti-necroptotic effects of glucocorticoids in human bronchial epithelial cells

  • Life Sci. 2019 Mar 15;221:168-177. doi: 10.1016/j.lfs.2019.02.013.
Huojun Zhang 1 Qian Liu 1 Luxia Kong 1 Shuyun Xu 2
Affiliations

Affiliations

  • 1 Department of Respiratory and Critical Care Medicine, Key Laboratory of Pulmonary Diseases of Health Ministry, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Hubei 430030, China.
  • 2 Department of Respiratory and Critical Care Medicine, Key Laboratory of Pulmonary Diseases of Health Ministry, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Hubei 430030, China. Electronic address: sxu@hust.edu.cn.
Abstract

Aims: To investigate whether Mucin 1 (MUC1) downregulation reduced the sensitivity of tumor necrosis factor-alpha (TNF-α)-induced bronchial epithelial cells to glucocorticoid-mediated Necroptosis and explore the underlying mechanisms.

Main methods: The human lung bronchial epithelial cell line (16HBE) was transfected with small interfering RNA (siRNA) against MUC1 and then stimulated by TNF-α, where some cells were pretreated with dexamethasone. Flow cytometry was performed to analyze Necroptosis in 16HBE cells, and western blot analysis was used to detect protein expression levels of MUC1, Glucocorticoid Receptor (GR)α, GRβ, NF-κB p65, phospho-p65 (p-p65), and histone deacetylase-2 (HDAC2). Additionally, nuclear translocation of MUC1 and GRα was assessed by immunofluorescence.

Key findings: We observed that MUC1 downregulation by siRNA significantly augmented TNF-α-induced Necroptosis in 16HBE cells, and that dexamethasone showed impaired anti-necroptotic effects of MUC1 downregulation. Furthermore, we found that GRα nuclear translocation was inhibited in 16HBE cells with MUC1 downregulation, and that dexamethasone-mediated inhibition of p65 phosphorylation was lower in cells transfected with MUC1-siRNA compared to those transfected with negative control siRNA.

Significance: Impaired GRα nuclear translocation and inhibited p-p65 expression might contribute to glucocorticoid resistance caused by MUC1 deficiency in TNF-α-induced Necroptosis in 16HBE cells, and should be considered as a potential target for the development of novel therapeutics for asthma.

Keywords

Glucocorticoid resistance; MUC1; Necroptosis; Nuclear translocation; P-p65.

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