Transcription Elongation Factor P-TEFb Is Involved in IL-17F Signaling in Airway Smooth Muscle Cells

Masayuki Nakajima, Mio Kawaguchi, Masashi Matsuyama, Kyoko Ota, Junichi Fujita, Satoshi Matsukura, Shau Ku Huang, Yuko Morishima, Yukio Ishii, Hiroaki Satoh, Tohru Sakamoto, Nobuyuki Hizawa

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: IL-17F is involved in the pathogenesis of several inflammatory diseases, including asthma and COPD. However, the effects of steroids on the function of IL-17F signaling mechanisms are largely unknown. One of the transcription elongation factors, positive transcription elongation factor b (P-TEFb) composed of cyclin T1 and cyclin-dependent kinase 9 (CDK9), is known as a novel checkpoint regulator of gene expression via bromodomain-containing protein 4 (Brd4). Methods: Human airway smooth muscle cells were stimulated with IL-17F and the expression of IL-8 was evaluated by real-time PCR and ELISA. Next, the phosphorylation of CDK9 was determined by Western blotting. The CDK9 inhibitor and short interfering RNAs (siRNAs) targeting Brd4, cyclin T1, and CDK9 were used to identify the effect on IL-17F-induced IL-8 expression. Finally, the effect of steroids and its signaling were evaluated. Results: IL-17F markedly induced the transcription of the IL-8 gene and the expression of the protein. Pretreatment of CDK9 inhibitor and transfection of siRNAs targeting CDK9 markedly abrogated IL-17F-induced IL-8 production. Transfection of siRNAs targeting Brd4 and cyclin T1 diminished IL-17F-induced phosphorylation of CDK9 and IL-8 production. Moreover, budesonide decreased CDK9 phosphorylation and markedly inhibited IL-17F-induced IL-8 production. Conclusions: This is the first report that P-TEFb is involved in IL-17F-induced IL-8 expression and that steroids diminish it via the inhibition of CDK9 phosphorylation. IL-17F and P-TEFb might be novel therapeutic targets for airway inflammatory diseases.

Original languageEnglish
JournalInternational Archives of Allergy and Immunology
DOIs
Publication statusAccepted/In press - Apr 12 2018
Externally publishedYes

Fingerprint

Cyclin-Dependent Kinase 9
Peptide Elongation Factors
Interleukin-17
Smooth Muscle Myocytes
Transcription Factors
Interleukin-8
Cyclin T
Phosphorylation
Small Interfering RNA
Steroids
Transfection
Proteins
Gene Expression
Budesonide
Regulator Genes
Chronic Obstructive Pulmonary Disease
Real-Time Polymerase Chain Reaction
Asthma

Keywords

  • Airway smooth muscle cells
  • Bromodomain-containing protein 4
  • Interleukin-17F
  • Interleukin-8
  • Positive transcription elongation factor b

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Transcription Elongation Factor P-TEFb Is Involved in IL-17F Signaling in Airway Smooth Muscle Cells. / Nakajima, Masayuki; Kawaguchi, Mio; Matsuyama, Masashi; Ota, Kyoko; Fujita, Junichi; Matsukura, Satoshi; Huang, Shau Ku; Morishima, Yuko; Ishii, Yukio; Satoh, Hiroaki; Sakamoto, Tohru; Hizawa, Nobuyuki.

In: International Archives of Allergy and Immunology, 12.04.2018.

Research output: Contribution to journalArticle

Nakajima, M, Kawaguchi, M, Matsuyama, M, Ota, K, Fujita, J, Matsukura, S, Huang, SK, Morishima, Y, Ishii, Y, Satoh, H, Sakamoto, T & Hizawa, N 2018, 'Transcription Elongation Factor P-TEFb Is Involved in IL-17F Signaling in Airway Smooth Muscle Cells', International Archives of Allergy and Immunology. https://doi.org/10.1159/000488154
Nakajima, Masayuki ; Kawaguchi, Mio ; Matsuyama, Masashi ; Ota, Kyoko ; Fujita, Junichi ; Matsukura, Satoshi ; Huang, Shau Ku ; Morishima, Yuko ; Ishii, Yukio ; Satoh, Hiroaki ; Sakamoto, Tohru ; Hizawa, Nobuyuki. / Transcription Elongation Factor P-TEFb Is Involved in IL-17F Signaling in Airway Smooth Muscle Cells. In: International Archives of Allergy and Immunology. 2018.
@article{6fc86698e86e45608ec322cdd04dc046,
title = "Transcription Elongation Factor P-TEFb Is Involved in IL-17F Signaling in Airway Smooth Muscle Cells",
abstract = "Background: IL-17F is involved in the pathogenesis of several inflammatory diseases, including asthma and COPD. However, the effects of steroids on the function of IL-17F signaling mechanisms are largely unknown. One of the transcription elongation factors, positive transcription elongation factor b (P-TEFb) composed of cyclin T1 and cyclin-dependent kinase 9 (CDK9), is known as a novel checkpoint regulator of gene expression via bromodomain-containing protein 4 (Brd4). Methods: Human airway smooth muscle cells were stimulated with IL-17F and the expression of IL-8 was evaluated by real-time PCR and ELISA. Next, the phosphorylation of CDK9 was determined by Western blotting. The CDK9 inhibitor and short interfering RNAs (siRNAs) targeting Brd4, cyclin T1, and CDK9 were used to identify the effect on IL-17F-induced IL-8 expression. Finally, the effect of steroids and its signaling were evaluated. Results: IL-17F markedly induced the transcription of the IL-8 gene and the expression of the protein. Pretreatment of CDK9 inhibitor and transfection of siRNAs targeting CDK9 markedly abrogated IL-17F-induced IL-8 production. Transfection of siRNAs targeting Brd4 and cyclin T1 diminished IL-17F-induced phosphorylation of CDK9 and IL-8 production. Moreover, budesonide decreased CDK9 phosphorylation and markedly inhibited IL-17F-induced IL-8 production. Conclusions: This is the first report that P-TEFb is involved in IL-17F-induced IL-8 expression and that steroids diminish it via the inhibition of CDK9 phosphorylation. IL-17F and P-TEFb might be novel therapeutic targets for airway inflammatory diseases.",
keywords = "Airway smooth muscle cells, Bromodomain-containing protein 4, Interleukin-17F, Interleukin-8, Positive transcription elongation factor b",
author = "Masayuki Nakajima and Mio Kawaguchi and Masashi Matsuyama and Kyoko Ota and Junichi Fujita and Satoshi Matsukura and Huang, {Shau Ku} and Yuko Morishima and Yukio Ishii and Hiroaki Satoh and Tohru Sakamoto and Nobuyuki Hizawa",
year = "2018",
month = "4",
day = "12",
doi = "10.1159/000488154",
language = "English",
journal = "International Archives of Allergy and Immunology",
issn = "1018-2438",
publisher = "S. Karger AG",

}

TY - JOUR

T1 - Transcription Elongation Factor P-TEFb Is Involved in IL-17F Signaling in Airway Smooth Muscle Cells

AU - Nakajima, Masayuki

AU - Kawaguchi, Mio

AU - Matsuyama, Masashi

AU - Ota, Kyoko

AU - Fujita, Junichi

AU - Matsukura, Satoshi

AU - Huang, Shau Ku

AU - Morishima, Yuko

AU - Ishii, Yukio

AU - Satoh, Hiroaki

AU - Sakamoto, Tohru

AU - Hizawa, Nobuyuki

PY - 2018/4/12

Y1 - 2018/4/12

N2 - Background: IL-17F is involved in the pathogenesis of several inflammatory diseases, including asthma and COPD. However, the effects of steroids on the function of IL-17F signaling mechanisms are largely unknown. One of the transcription elongation factors, positive transcription elongation factor b (P-TEFb) composed of cyclin T1 and cyclin-dependent kinase 9 (CDK9), is known as a novel checkpoint regulator of gene expression via bromodomain-containing protein 4 (Brd4). Methods: Human airway smooth muscle cells were stimulated with IL-17F and the expression of IL-8 was evaluated by real-time PCR and ELISA. Next, the phosphorylation of CDK9 was determined by Western blotting. The CDK9 inhibitor and short interfering RNAs (siRNAs) targeting Brd4, cyclin T1, and CDK9 were used to identify the effect on IL-17F-induced IL-8 expression. Finally, the effect of steroids and its signaling were evaluated. Results: IL-17F markedly induced the transcription of the IL-8 gene and the expression of the protein. Pretreatment of CDK9 inhibitor and transfection of siRNAs targeting CDK9 markedly abrogated IL-17F-induced IL-8 production. Transfection of siRNAs targeting Brd4 and cyclin T1 diminished IL-17F-induced phosphorylation of CDK9 and IL-8 production. Moreover, budesonide decreased CDK9 phosphorylation and markedly inhibited IL-17F-induced IL-8 production. Conclusions: This is the first report that P-TEFb is involved in IL-17F-induced IL-8 expression and that steroids diminish it via the inhibition of CDK9 phosphorylation. IL-17F and P-TEFb might be novel therapeutic targets for airway inflammatory diseases.

AB - Background: IL-17F is involved in the pathogenesis of several inflammatory diseases, including asthma and COPD. However, the effects of steroids on the function of IL-17F signaling mechanisms are largely unknown. One of the transcription elongation factors, positive transcription elongation factor b (P-TEFb) composed of cyclin T1 and cyclin-dependent kinase 9 (CDK9), is known as a novel checkpoint regulator of gene expression via bromodomain-containing protein 4 (Brd4). Methods: Human airway smooth muscle cells were stimulated with IL-17F and the expression of IL-8 was evaluated by real-time PCR and ELISA. Next, the phosphorylation of CDK9 was determined by Western blotting. The CDK9 inhibitor and short interfering RNAs (siRNAs) targeting Brd4, cyclin T1, and CDK9 were used to identify the effect on IL-17F-induced IL-8 expression. Finally, the effect of steroids and its signaling were evaluated. Results: IL-17F markedly induced the transcription of the IL-8 gene and the expression of the protein. Pretreatment of CDK9 inhibitor and transfection of siRNAs targeting CDK9 markedly abrogated IL-17F-induced IL-8 production. Transfection of siRNAs targeting Brd4 and cyclin T1 diminished IL-17F-induced phosphorylation of CDK9 and IL-8 production. Moreover, budesonide decreased CDK9 phosphorylation and markedly inhibited IL-17F-induced IL-8 production. Conclusions: This is the first report that P-TEFb is involved in IL-17F-induced IL-8 expression and that steroids diminish it via the inhibition of CDK9 phosphorylation. IL-17F and P-TEFb might be novel therapeutic targets for airway inflammatory diseases.

KW - Airway smooth muscle cells

KW - Bromodomain-containing protein 4

KW - Interleukin-17F

KW - Interleukin-8

KW - Positive transcription elongation factor b

UR - http://www.scopus.com/inward/record.url?scp=85045303672&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85045303672&partnerID=8YFLogxK

U2 - 10.1159/000488154

DO - 10.1159/000488154

M3 - Article

JO - International Archives of Allergy and Immunology

JF - International Archives of Allergy and Immunology

SN - 1018-2438

ER -