Phorbol 12-myristate 13-acetate upregulates cyclooxygenase-2 expression in human pulmonary epithelial cells via Ras, Raf-1, ERK, and NF-κB, but not p38 MAPK, pathways

Ming Shyan Chang, Bing Chang Chen, Ming Tze Yu, Joen Rong Sheu, Tseng Fu Chen, Chien Huang Lin

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Abstract

In this study, we investigated the signaling pathway involved in cyclooxygenase-2 (COX-2) expression and prostaglandin E 2 (PGE 2) release by phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, in human pulmonary epithelial cells (A549). PMA-induced COX-2 expression was attenuated by PKC inhibitors (Go 6976 and Ro 31-8220), a Ras inhibitor (manumycin A), a Raf-1 inhibitor (GW 5074), a MEK inhibitor (PD 098059), and an NF-κB inhibitor (PDTC), but not by a tyrosine kinase inhibitor (genistein) or a p38 MAPK inhibitor (SB 203580). PMA also caused the activation of Ras, Raf-1, and ERK1/2. PMA-induced activation of Ras and Raf-1 was inhibited by Ro 31-8220 and manumycin A. PMA-mediated activation of ERK1/2 was inhibited by Ro 31-8220, manumycin A, GW 5074, and PD 098059. Stimulation of cells with PMA caused IκBα phosphorylation, IκBα degradation, and the formation of a NF-κB-specific DNA-protein complex. The PMA-mediated increase in κB-luciferase activity was inhibited by Ro 31-8220, manumycin A, GW5074, PD 098059, and PDTC. Taken together, these results indicate that PMA might activate PKC to elicit activation of the Ras/Raf-1/ERK1/2 pathway, which in turn initiates NF-κB activation, and finally induces COX-2 expression and PGE 2 release in A549 cells.

Original languageEnglish
Pages (from-to)299-310
Number of pages12
JournalCellular Signalling
Volume17
Issue number3
DOIs
Publication statusPublished - Mar 2005
Externally publishedYes

Fingerprint

p38 Mitogen-Activated Protein Kinases
Cyclooxygenase 2
Acetates
Up-Regulation
Epithelial Cells
Lung
Protein Kinase C
Prostaglandins E
phorbol-12-myristate
MAP Kinase Signaling System
Protein C Inhibitor
Genistein
Mitogen-Activated Protein Kinase Kinases
Protein Kinase Inhibitors
Luciferases
Protein-Tyrosine Kinases
Phosphorylation
Ro 31-8220
manumycin
DNA

Keywords

  • A549 cells
  • Cyclooxygenase-2
  • ERK1/2
  • NF-κB
  • PKC
  • PMA
  • Raf-1
  • Ras

ASJC Scopus subject areas

  • Cell Biology

Cite this

@article{e3d16b82d9d640a0ae8d1c78b0802f58,
title = "Phorbol 12-myristate 13-acetate upregulates cyclooxygenase-2 expression in human pulmonary epithelial cells via Ras, Raf-1, ERK, and NF-κB, but not p38 MAPK, pathways",
abstract = "In this study, we investigated the signaling pathway involved in cyclooxygenase-2 (COX-2) expression and prostaglandin E 2 (PGE 2) release by phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, in human pulmonary epithelial cells (A549). PMA-induced COX-2 expression was attenuated by PKC inhibitors (Go 6976 and Ro 31-8220), a Ras inhibitor (manumycin A), a Raf-1 inhibitor (GW 5074), a MEK inhibitor (PD 098059), and an NF-κB inhibitor (PDTC), but not by a tyrosine kinase inhibitor (genistein) or a p38 MAPK inhibitor (SB 203580). PMA also caused the activation of Ras, Raf-1, and ERK1/2. PMA-induced activation of Ras and Raf-1 was inhibited by Ro 31-8220 and manumycin A. PMA-mediated activation of ERK1/2 was inhibited by Ro 31-8220, manumycin A, GW 5074, and PD 098059. Stimulation of cells with PMA caused IκBα phosphorylation, IκBα degradation, and the formation of a NF-κB-specific DNA-protein complex. The PMA-mediated increase in κB-luciferase activity was inhibited by Ro 31-8220, manumycin A, GW5074, PD 098059, and PDTC. Taken together, these results indicate that PMA might activate PKC to elicit activation of the Ras/Raf-1/ERK1/2 pathway, which in turn initiates NF-κB activation, and finally induces COX-2 expression and PGE 2 release in A549 cells.",
keywords = "A549 cells, Cyclooxygenase-2, ERK1/2, NF-κB, PKC, PMA, Raf-1, Ras",
author = "Chang, {Ming Shyan} and Chen, {Bing Chang} and Yu, {Ming Tze} and Sheu, {Joen Rong} and Chen, {Tseng Fu} and Lin, {Chien Huang}",
year = "2005",
month = "3",
doi = "10.1016/j.cellsig.2004.07.008",
language = "English",
volume = "17",
pages = "299--310",
journal = "Cellular Signalling",
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T1 - Phorbol 12-myristate 13-acetate upregulates cyclooxygenase-2 expression in human pulmonary epithelial cells via Ras, Raf-1, ERK, and NF-κB, but not p38 MAPK, pathways

AU - Chang, Ming Shyan

AU - Chen, Bing Chang

AU - Yu, Ming Tze

AU - Sheu, Joen Rong

AU - Chen, Tseng Fu

AU - Lin, Chien Huang

PY - 2005/3

Y1 - 2005/3

N2 - In this study, we investigated the signaling pathway involved in cyclooxygenase-2 (COX-2) expression and prostaglandin E 2 (PGE 2) release by phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, in human pulmonary epithelial cells (A549). PMA-induced COX-2 expression was attenuated by PKC inhibitors (Go 6976 and Ro 31-8220), a Ras inhibitor (manumycin A), a Raf-1 inhibitor (GW 5074), a MEK inhibitor (PD 098059), and an NF-κB inhibitor (PDTC), but not by a tyrosine kinase inhibitor (genistein) or a p38 MAPK inhibitor (SB 203580). PMA also caused the activation of Ras, Raf-1, and ERK1/2. PMA-induced activation of Ras and Raf-1 was inhibited by Ro 31-8220 and manumycin A. PMA-mediated activation of ERK1/2 was inhibited by Ro 31-8220, manumycin A, GW 5074, and PD 098059. Stimulation of cells with PMA caused IκBα phosphorylation, IκBα degradation, and the formation of a NF-κB-specific DNA-protein complex. The PMA-mediated increase in κB-luciferase activity was inhibited by Ro 31-8220, manumycin A, GW5074, PD 098059, and PDTC. Taken together, these results indicate that PMA might activate PKC to elicit activation of the Ras/Raf-1/ERK1/2 pathway, which in turn initiates NF-κB activation, and finally induces COX-2 expression and PGE 2 release in A549 cells.

AB - In this study, we investigated the signaling pathway involved in cyclooxygenase-2 (COX-2) expression and prostaglandin E 2 (PGE 2) release by phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, in human pulmonary epithelial cells (A549). PMA-induced COX-2 expression was attenuated by PKC inhibitors (Go 6976 and Ro 31-8220), a Ras inhibitor (manumycin A), a Raf-1 inhibitor (GW 5074), a MEK inhibitor (PD 098059), and an NF-κB inhibitor (PDTC), but not by a tyrosine kinase inhibitor (genistein) or a p38 MAPK inhibitor (SB 203580). PMA also caused the activation of Ras, Raf-1, and ERK1/2. PMA-induced activation of Ras and Raf-1 was inhibited by Ro 31-8220 and manumycin A. PMA-mediated activation of ERK1/2 was inhibited by Ro 31-8220, manumycin A, GW 5074, and PD 098059. Stimulation of cells with PMA caused IκBα phosphorylation, IκBα degradation, and the formation of a NF-κB-specific DNA-protein complex. The PMA-mediated increase in κB-luciferase activity was inhibited by Ro 31-8220, manumycin A, GW5074, PD 098059, and PDTC. Taken together, these results indicate that PMA might activate PKC to elicit activation of the Ras/Raf-1/ERK1/2 pathway, which in turn initiates NF-κB activation, and finally induces COX-2 expression and PGE 2 release in A549 cells.

KW - A549 cells

KW - Cyclooxygenase-2

KW - ERK1/2

KW - NF-κB

KW - PKC

KW - PMA

KW - Raf-1

KW - Ras

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U2 - 10.1016/j.cellsig.2004.07.008

DO - 10.1016/j.cellsig.2004.07.008

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SN - 0898-6568

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