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Molecular Pharmacology

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Research ArticleArticle

Involvement of Protein Kinase C-γ in IL-1β-Induced Cyclooxygenase-2 Expression in Human Pulmonary Epithelial Cells

Chien-Huang Lin, Sheng-Yuan Sheu, Horng-Mo Lee, Yuan-Soon Ho, Wen-Sen Lee, Wun-Chang Ko and Joen-Rong Sheu
Molecular Pharmacology January 2000, 57 (1) 36-43;
Chien-Huang Lin
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Sheng-Yuan Sheu
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Horng-Mo Lee
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Yuan-Soon Ho
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Wen-Sen Lee
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Wun-Chang Ko
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Joen-Rong Sheu
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Abstract

The signaling pathway of protein kinase C (PKC) is known to play a role in mediating the action of various cytokines. Here we examined the signal transduction pathway of PKC activation and the role of PKC isoforms in interleukin-1β (IL-1β)-mediated cyclooxygenase-2 (COX-2) expression in human pulmonary epithelial cell line (A549). The tyrosine kinase inhibitors (genistein and tyrphostin AG126) and phosphatidylcholine-phospholipase C inhibitor (D-609) prevented IL-1β-induced prostaglandin E2 (PGE2) release and COX-2 expression, whereas U-73122 (a phosphatidylinositol-phospholipase C inhibitor) and propranolol (a phosphatidate phosphohydrolase inhibitor) had no effect. The PKC inhibitors (Go 6976 and Ro 31–8220) and NF-κB inhibitor, pyrrolidine dithiocarbamate, also attenuated IL-1β-induced PGE2release and COX-2 expression. Western blot analysis using PKC isoenzyme-specific antibodies indicated that A549 cells expressed PKC-α, -γ, -ι, -λ, -ζ, and -μ. IL-1β caused the translocation of PKC-γ but not other isoforms from cytosol to the membrane fraction. Moreover, the translocation of PKC-γ was inhibited by genistein or D-609, but not by U-73122. IL-1β caused the translocation of p65 NF-κB from cytosol to the nucleus as well as the degradation of IκB-α in cytosol. Furthermore, the translocation of p65 NF-κB was inhibited by genistein, Go 6976, Ro 31–8220, or pyrrolidine dithiocarbamate. These results indicate that in human pulmonary epithelial cells, IL-1β might activate phosphatidylcholine-phospholipase C through an upstream tyrosine phosphorylation to elicit PKC activation, which in turn initiates NF-κB activation, and finally induces COX-2 expression and PGE2 release. Of the PKC isoforms present in A549 cells, only activation of PKC-γ is involved in regulating IL-1β-induced responses.

Footnotes

  • Send reprint requests to: Chien-Huang Lin, Ph.D., Institute of Biomedical Technology, Taipei Medical College, 250 Wu-Hsing St., Taipei 110, Taiwan. E-mail: chlin{at}tmc.edu.tw

  • This work was supported by the National Science Council of the Republic of China Research Grants NSC87–2314-B-038–052 and NSC88–2314-B-038–131.

  • Abbreviations:
    COX
    cyclooxygenase
    IL-1β
    interleukin-1β
    PGE2
    prostaglandin E2
    PC-PLC
    phosphatidylcholine-phospholipase C
    PI-PLC
    phosphatidylinositol-phospholipase C
    DAG
    diacylglycerol
    PKC
    protein kinase C
    PDTC
    pyrrolidine dithiocarbamate
    DMEM
    Dulbecco's modified Eagle's medium
    FCS
    fetal calf serum
    DTT
    dithiothreitol
    PMSF
    phenylmethylsulfonyl fluoride
    NP-40
    Nonident P-40
    • Received May 24, 1999.
    • Accepted October 10, 1999.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 57 (1)
Molecular Pharmacology
Vol. 57, Issue 1
1 Jan 2000
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Research ArticleArticle

Involvement of Protein Kinase C-γ in IL-1β-Induced Cyclooxygenase-2 Expression in Human Pulmonary Epithelial Cells

Chien-Huang Lin, Sheng-Yuan Sheu, Horng-Mo Lee, Yuan-Soon Ho, Wen-Sen Lee, Wun-Chang Ko and Joen-Rong Sheu
Molecular Pharmacology January 1, 2000, 57 (1) 36-43;

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Research ArticleArticle

Involvement of Protein Kinase C-γ in IL-1β-Induced Cyclooxygenase-2 Expression in Human Pulmonary Epithelial Cells

Chien-Huang Lin, Sheng-Yuan Sheu, Horng-Mo Lee, Yuan-Soon Ho, Wen-Sen Lee, Wun-Chang Ko and Joen-Rong Sheu
Molecular Pharmacology January 1, 2000, 57 (1) 36-43;
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