Synthetic double-stranded RNA induces interleukin-32 in bronchial epithelial cells

Kyoko Ota, Mio Kawaguchi, Junichi Fujita, Fumio Kokubu, Shau Ku Huang, Yuko Morishima, Satoshi Matsukura, Masatsugu Kurokawa, Yukio Ishii, Hiroaki Satoh, Tohru Sakamoto, Nobuyuki Hizawa

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Objective: Interleukin (IL)-32 is a novel cytokine and is involved in the pathogenesis of various inflammatory diseases, including asthma and COPD. However, the regulatory mechanisms of IL-32 expression and its precise pathogenic role remain to be defined. Given that viral infections are known to potentially cause and exacerbate airway inflammation, in this study, we investigated the expression of IL-32 induced by synthetic double-stranded (ds) RNA, and its signaling mechanisms involved. Methods: Bronchial epithelial cells were stimulated with synthetic dsRNA poly I:C. The levels of IL-32 expression were analyzed using real-time PCR and ELISA. The involvement of transforming growth factor β-activated kinase 1 (TAK1) and a subunit of nuclear factor-κB (NF-κB), p65 was determined by western blot analyses. TAK1 inhibitor, 5Z-7-Oxozeaenol and NF-κB inhibitor, BAY 11-7082 were added to the culture to identify key signaling events leading to the expression of IL-32. Finally, the effect of short interfering RNAs (siRNAs) targeting TAK1 and p65 was investigated. Results: dsRNA significantly induced IL-32 gene and protein expression, concomitant with activation of TAK1 and p65. Pretreatment of 5Z-7-Oxozeaenol diminished dsRNA-induced phosphorylation of NF-κB. Both 5Z-7-Oxozeaenol and BAY 11-7082 significantly abrogated dsRNA-induced IL-32 production. Moreover, transfection of the cells with siRNAs targeting TAK1 and p65 inhibited the expression of IL-32. Conclusions: The expression of IL-32 is induced by dsRNA via the TAK1-NF-κB signaling pathway in bronchial epithelial cells. IL-32 is involved in the pathogenesis of airway inflammation, and may be a novel therapeutic target for airway inflammatory diseases.

Original languageEnglish
Pages (from-to)335-343
Number of pages9
JournalExperimental Lung Research
Volume41
Issue number6
DOIs
Publication statusPublished - Jan 1 2015
Externally publishedYes

Fingerprint

Double-Stranded RNA
Interleukins
Epithelial Cells
Small Interfering RNA
Inflammation
Poly I-C
Phosphorylation
Transforming Growth Factors
Virus Diseases
Chronic Obstructive Pulmonary Disease
Transfection
Real-Time Polymerase Chain Reaction
Phosphotransferases
Asthma
Western Blotting
Chemical activation
Enzyme-Linked Immunosorbent Assay
Cytokines
Gene Expression

Keywords

  • bronchial epithelial cell
  • dsRNA
  • IL-32
  • NF-κB
  • p65
  • TAK1

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry

Cite this

Synthetic double-stranded RNA induces interleukin-32 in bronchial epithelial cells. / Ota, Kyoko; Kawaguchi, Mio; Fujita, Junichi; Kokubu, Fumio; Huang, Shau Ku; Morishima, Yuko; Matsukura, Satoshi; Kurokawa, Masatsugu; Ishii, Yukio; Satoh, Hiroaki; Sakamoto, Tohru; Hizawa, Nobuyuki.

In: Experimental Lung Research, Vol. 41, No. 6, 01.01.2015, p. 335-343.

Research output: Contribution to journalArticle

Ota, K, Kawaguchi, M, Fujita, J, Kokubu, F, Huang, SK, Morishima, Y, Matsukura, S, Kurokawa, M, Ishii, Y, Satoh, H, Sakamoto, T & Hizawa, N 2015, 'Synthetic double-stranded RNA induces interleukin-32 in bronchial epithelial cells', Experimental Lung Research, vol. 41, no. 6, pp. 335-343. https://doi.org/10.3109/01902148.2015.1033569
Ota, Kyoko ; Kawaguchi, Mio ; Fujita, Junichi ; Kokubu, Fumio ; Huang, Shau Ku ; Morishima, Yuko ; Matsukura, Satoshi ; Kurokawa, Masatsugu ; Ishii, Yukio ; Satoh, Hiroaki ; Sakamoto, Tohru ; Hizawa, Nobuyuki. / Synthetic double-stranded RNA induces interleukin-32 in bronchial epithelial cells. In: Experimental Lung Research. 2015 ; Vol. 41, No. 6. pp. 335-343.
@article{bd1932fe199a465fbd6207fdf2e96ea3,
title = "Synthetic double-stranded RNA induces interleukin-32 in bronchial epithelial cells",
abstract = "Objective: Interleukin (IL)-32 is a novel cytokine and is involved in the pathogenesis of various inflammatory diseases, including asthma and COPD. However, the regulatory mechanisms of IL-32 expression and its precise pathogenic role remain to be defined. Given that viral infections are known to potentially cause and exacerbate airway inflammation, in this study, we investigated the expression of IL-32 induced by synthetic double-stranded (ds) RNA, and its signaling mechanisms involved. Methods: Bronchial epithelial cells were stimulated with synthetic dsRNA poly I:C. The levels of IL-32 expression were analyzed using real-time PCR and ELISA. The involvement of transforming growth factor β-activated kinase 1 (TAK1) and a subunit of nuclear factor-κB (NF-κB), p65 was determined by western blot analyses. TAK1 inhibitor, 5Z-7-Oxozeaenol and NF-κB inhibitor, BAY 11-7082 were added to the culture to identify key signaling events leading to the expression of IL-32. Finally, the effect of short interfering RNAs (siRNAs) targeting TAK1 and p65 was investigated. Results: dsRNA significantly induced IL-32 gene and protein expression, concomitant with activation of TAK1 and p65. Pretreatment of 5Z-7-Oxozeaenol diminished dsRNA-induced phosphorylation of NF-κB. Both 5Z-7-Oxozeaenol and BAY 11-7082 significantly abrogated dsRNA-induced IL-32 production. Moreover, transfection of the cells with siRNAs targeting TAK1 and p65 inhibited the expression of IL-32. Conclusions: The expression of IL-32 is induced by dsRNA via the TAK1-NF-κB signaling pathway in bronchial epithelial cells. IL-32 is involved in the pathogenesis of airway inflammation, and may be a novel therapeutic target for airway inflammatory diseases.",
keywords = "bronchial epithelial cell, dsRNA, IL-32, NF-κB, p65, TAK1",
author = "Kyoko Ota and Mio Kawaguchi and Junichi Fujita and Fumio Kokubu and Huang, {Shau Ku} and Yuko Morishima and Satoshi Matsukura and Masatsugu Kurokawa and Yukio Ishii and Hiroaki Satoh and Tohru Sakamoto and Nobuyuki Hizawa",
year = "2015",
month = "1",
day = "1",
doi = "10.3109/01902148.2015.1033569",
language = "English",
volume = "41",
pages = "335--343",
journal = "Experimental Lung Research",
issn = "0190-2148",
publisher = "Informa Healthcare",
number = "6",

}

TY - JOUR

T1 - Synthetic double-stranded RNA induces interleukin-32 in bronchial epithelial cells

AU - Ota, Kyoko

AU - Kawaguchi, Mio

AU - Fujita, Junichi

AU - Kokubu, Fumio

AU - Huang, Shau Ku

AU - Morishima, Yuko

AU - Matsukura, Satoshi

AU - Kurokawa, Masatsugu

AU - Ishii, Yukio

AU - Satoh, Hiroaki

AU - Sakamoto, Tohru

AU - Hizawa, Nobuyuki

PY - 2015/1/1

Y1 - 2015/1/1

N2 - Objective: Interleukin (IL)-32 is a novel cytokine and is involved in the pathogenesis of various inflammatory diseases, including asthma and COPD. However, the regulatory mechanisms of IL-32 expression and its precise pathogenic role remain to be defined. Given that viral infections are known to potentially cause and exacerbate airway inflammation, in this study, we investigated the expression of IL-32 induced by synthetic double-stranded (ds) RNA, and its signaling mechanisms involved. Methods: Bronchial epithelial cells were stimulated with synthetic dsRNA poly I:C. The levels of IL-32 expression were analyzed using real-time PCR and ELISA. The involvement of transforming growth factor β-activated kinase 1 (TAK1) and a subunit of nuclear factor-κB (NF-κB), p65 was determined by western blot analyses. TAK1 inhibitor, 5Z-7-Oxozeaenol and NF-κB inhibitor, BAY 11-7082 were added to the culture to identify key signaling events leading to the expression of IL-32. Finally, the effect of short interfering RNAs (siRNAs) targeting TAK1 and p65 was investigated. Results: dsRNA significantly induced IL-32 gene and protein expression, concomitant with activation of TAK1 and p65. Pretreatment of 5Z-7-Oxozeaenol diminished dsRNA-induced phosphorylation of NF-κB. Both 5Z-7-Oxozeaenol and BAY 11-7082 significantly abrogated dsRNA-induced IL-32 production. Moreover, transfection of the cells with siRNAs targeting TAK1 and p65 inhibited the expression of IL-32. Conclusions: The expression of IL-32 is induced by dsRNA via the TAK1-NF-κB signaling pathway in bronchial epithelial cells. IL-32 is involved in the pathogenesis of airway inflammation, and may be a novel therapeutic target for airway inflammatory diseases.

AB - Objective: Interleukin (IL)-32 is a novel cytokine and is involved in the pathogenesis of various inflammatory diseases, including asthma and COPD. However, the regulatory mechanisms of IL-32 expression and its precise pathogenic role remain to be defined. Given that viral infections are known to potentially cause and exacerbate airway inflammation, in this study, we investigated the expression of IL-32 induced by synthetic double-stranded (ds) RNA, and its signaling mechanisms involved. Methods: Bronchial epithelial cells were stimulated with synthetic dsRNA poly I:C. The levels of IL-32 expression were analyzed using real-time PCR and ELISA. The involvement of transforming growth factor β-activated kinase 1 (TAK1) and a subunit of nuclear factor-κB (NF-κB), p65 was determined by western blot analyses. TAK1 inhibitor, 5Z-7-Oxozeaenol and NF-κB inhibitor, BAY 11-7082 were added to the culture to identify key signaling events leading to the expression of IL-32. Finally, the effect of short interfering RNAs (siRNAs) targeting TAK1 and p65 was investigated. Results: dsRNA significantly induced IL-32 gene and protein expression, concomitant with activation of TAK1 and p65. Pretreatment of 5Z-7-Oxozeaenol diminished dsRNA-induced phosphorylation of NF-κB. Both 5Z-7-Oxozeaenol and BAY 11-7082 significantly abrogated dsRNA-induced IL-32 production. Moreover, transfection of the cells with siRNAs targeting TAK1 and p65 inhibited the expression of IL-32. Conclusions: The expression of IL-32 is induced by dsRNA via the TAK1-NF-κB signaling pathway in bronchial epithelial cells. IL-32 is involved in the pathogenesis of airway inflammation, and may be a novel therapeutic target for airway inflammatory diseases.

KW - bronchial epithelial cell

KW - dsRNA

KW - IL-32

KW - NF-κB

KW - p65

KW - TAK1

UR - http://www.scopus.com/inward/record.url?scp=84938360982&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84938360982&partnerID=8YFLogxK

U2 - 10.3109/01902148.2015.1033569

DO - 10.3109/01902148.2015.1033569

M3 - Article

VL - 41

SP - 335

EP - 343

JO - Experimental Lung Research

JF - Experimental Lung Research

SN - 0190-2148

IS - 6

ER -