NF-κB mediates IL-1β- and IL-17A-induced MUC5B expression in airway epithelial cells

Tomoyuki Fujisawa, Mary Mann Jong Chang, Sharlene Velichko, Philip Thai, Li Yin Hung, Fei Huang, Newton Phuong, Yin Chen, Reen Wu

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

A major pathological feature of chronic airway diseases is the elevated expression of gel-forming mucins. NF-κB activation in airway epithelial cells has been shown to play a proinflammatory role in chronic airway diseases; however, the specific role of NF-κB in mucin gene expression has not been characterized. In this study, we show that the proinflammatory cytokines, IL-1β and IL-17A, both of which use the NF-κB pathway, are potent inducers of MUC5B mRNA expression in both well differentiated primary normal human bronchial epithelial cells and the human bronchial epithelial cell line, HBE1. MUC5B induction by these cytokines was both time- and dose-dependent, and was attenuated by the small molecule inhibitor, NF-κB inhibitor III, as well as p65 small interfering RNA, suggesting that the regulation of MUC5B expression by these cytokines is via an NF-κB-based transcriptional mechanism. Deletion analysis of the MUC5B promoter demonstrated that IL-1β- and IL-17A-induced promoter activity resides within the -4.17-kb to -2.56-kb region relative to the transcriptional start site. This region contains three putative κB-binding sites (NF-κB-1, -3,786/-3,774; NF-κB-2, -3,173/-3,161; and NF-κB-3, -2,921/-2,909). Chromatin immunoprecipitation analysis confirmed enhanced binding of the p50 NF-κB subunit to the NF-κB-3 site after cytokine stimulation. We conclude that an NF-κB-based transcriptional mechanism is involved in MUC5B regulation by IL-1β and IL-17A in airway epithelium. This is the first demonstration of the participation of NF-κB and its specific binding site in cytokine-mediated airway MUC5B expression.

Original languageEnglish (US)
Pages (from-to)246-252
Number of pages7
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume45
Issue number2
DOIs
StatePublished - Aug 1 2011

Fingerprint

Interleukin-17
Interleukin-1
Epithelial Cells
Cytokines
Mucins
Chronic Disease
Binding Sites
Chromatin Immunoprecipitation
Gene expression
Small Interfering RNA
Chromatin
Demonstrations
Epithelium
Gels
Chemical activation
Gene Expression
Cell Line
Messenger RNA
Molecules

Keywords

  • Cytokines
  • Gene regulation
  • Lung
  • Mucin
  • Transcription factors

ASJC Scopus subject areas

  • Cell Biology
  • Pulmonary and Respiratory Medicine
  • Molecular Biology
  • Clinical Biochemistry

Cite this

NF-κB mediates IL-1β- and IL-17A-induced MUC5B expression in airway epithelial cells. / Fujisawa, Tomoyuki; Chang, Mary Mann Jong; Velichko, Sharlene; Thai, Philip; Hung, Li Yin; Huang, Fei; Phuong, Newton; Chen, Yin; Wu, Reen.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 45, No. 2, 01.08.2011, p. 246-252.

Research output: Contribution to journalArticle

Fujisawa, Tomoyuki ; Chang, Mary Mann Jong ; Velichko, Sharlene ; Thai, Philip ; Hung, Li Yin ; Huang, Fei ; Phuong, Newton ; Chen, Yin ; Wu, Reen. / NF-κB mediates IL-1β- and IL-17A-induced MUC5B expression in airway epithelial cells. In: American Journal of Respiratory Cell and Molecular Biology. 2011 ; Vol. 45, No. 2. pp. 246-252.
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abstract = "A major pathological feature of chronic airway diseases is the elevated expression of gel-forming mucins. NF-κB activation in airway epithelial cells has been shown to play a proinflammatory role in chronic airway diseases; however, the specific role of NF-κB in mucin gene expression has not been characterized. In this study, we show that the proinflammatory cytokines, IL-1β and IL-17A, both of which use the NF-κB pathway, are potent inducers of MUC5B mRNA expression in both well differentiated primary normal human bronchial epithelial cells and the human bronchial epithelial cell line, HBE1. MUC5B induction by these cytokines was both time- and dose-dependent, and was attenuated by the small molecule inhibitor, NF-κB inhibitor III, as well as p65 small interfering RNA, suggesting that the regulation of MUC5B expression by these cytokines is via an NF-κB-based transcriptional mechanism. Deletion analysis of the MUC5B promoter demonstrated that IL-1β- and IL-17A-induced promoter activity resides within the -4.17-kb to -2.56-kb region relative to the transcriptional start site. This region contains three putative κB-binding sites (NF-κB-1, -3,786/-3,774; NF-κB-2, -3,173/-3,161; and NF-κB-3, -2,921/-2,909). Chromatin immunoprecipitation analysis confirmed enhanced binding of the p50 NF-κB subunit to the NF-κB-3 site after cytokine stimulation. We conclude that an NF-κB-based transcriptional mechanism is involved in MUC5B regulation by IL-1β and IL-17A in airway epithelium. This is the first demonstration of the participation of NF-κB and its specific binding site in cytokine-mediated airway MUC5B expression.",
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AU - Fujisawa, Tomoyuki

AU - Chang, Mary Mann Jong

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AU - Thai, Philip

AU - Hung, Li Yin

AU - Huang, Fei

AU - Phuong, Newton

AU - Chen, Yin

AU - Wu, Reen

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AB - A major pathological feature of chronic airway diseases is the elevated expression of gel-forming mucins. NF-κB activation in airway epithelial cells has been shown to play a proinflammatory role in chronic airway diseases; however, the specific role of NF-κB in mucin gene expression has not been characterized. In this study, we show that the proinflammatory cytokines, IL-1β and IL-17A, both of which use the NF-κB pathway, are potent inducers of MUC5B mRNA expression in both well differentiated primary normal human bronchial epithelial cells and the human bronchial epithelial cell line, HBE1. MUC5B induction by these cytokines was both time- and dose-dependent, and was attenuated by the small molecule inhibitor, NF-κB inhibitor III, as well as p65 small interfering RNA, suggesting that the regulation of MUC5B expression by these cytokines is via an NF-κB-based transcriptional mechanism. Deletion analysis of the MUC5B promoter demonstrated that IL-1β- and IL-17A-induced promoter activity resides within the -4.17-kb to -2.56-kb region relative to the transcriptional start site. This region contains three putative κB-binding sites (NF-κB-1, -3,786/-3,774; NF-κB-2, -3,173/-3,161; and NF-κB-3, -2,921/-2,909). Chromatin immunoprecipitation analysis confirmed enhanced binding of the p50 NF-κB subunit to the NF-κB-3 site after cytokine stimulation. We conclude that an NF-κB-based transcriptional mechanism is involved in MUC5B regulation by IL-1β and IL-17A in airway epithelium. This is the first demonstration of the participation of NF-κB and its specific binding site in cytokine-mediated airway MUC5B expression.

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