Abstract

Our previous study demonstrated that 3-(5′-hydroxymethyl-2′-furyl)-1-benzylindazole (YC-1) might activate the soluble guanylate cyclase (sGC)/cGMP/protein kinase G (PKG) pathway to induce cyclooxygenase-2 (COX-2) expression in human pulmonary epithelial cells (A549). In this study, we further investigated the role of Raf-1 in YC-1-induced nuclear factor-κB (NF-κB) activation and COX-2 expression in A549 cells. YC-1-induced COX-2 expression was attenuated by a Raf-1 inhibitor (GW 5074) in a concentration-dependent manner. Treatment of A549 cells with YC-1 or 8-bromo-cGMP, a cell-permeable cGMP analogue, induced Raf-1 Ser338 phosphorylation in a time-dependent manner. YC-1-mediated Raf-1 activation was inhibited by an sGC inhibitor (ODQ), a PKG inhibitor (KT-5823), a Ras inhibitor (manumycin A), a dominant negative Ras mutant (RasN17), a protein kinase C-α (PKC-α) inhibitor (Ro 32-0432), and a phosphoinositide-3-OH-kinase (PI3K) inhibitor (LY 294002). Pretreatment of A549 cells with either manumycin A or GW 5074 attenuated YC-1-induced p44/42 MAPK activation. The YC-1-mediated increase in IKKα/β activation and κB-luciferase activity were attenuated by GW 5074, a MAPK/ERK kinase (MEK) inhibitor (PD 98059), and an ERK2 inhibitor (AG 126). Furthermore, YC-1-induced COX-2 promoter activity was also inhibited by GW 5074, PD 98059, and AG 126. These results indicate that YC-1 might activate the sGC/cGMP/PKG pathway to elicit Ras/Raf-1/p44/42 MAPK activation, which in turn induces IKKα/β and NF-κB activation, and ultimately causes COX-2 expression in A549 cells. Moreover, PKC-α and PI3K signal might be involved in YC-1-induced Raf-1 activation.

Original languageEnglish
Pages (from-to)247-253
Number of pages7
JournalPharmacological Research
Volume60
Issue number4
DOIs
Publication statusPublished - Oct 2009

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Mitogen-Activated Protein Kinase 3
Cyclooxygenase 2
Cyclic GMP-Dependent Protein Kinases
Epithelial Cells
Lung
Protein Kinase Inhibitors
Phosphatidylinositols
Protein Kinase C
Phosphotransferases
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Protein C Inhibitor
Mitogen-Activated Protein Kinase Kinases
Luciferases
Phosphorylation
A549 Cells
5-iodo-3-((3,5-dibromo-4-hydroxyphenyl)methylene)-2-indolinone
Soluble Guanylyl Cyclase

Keywords

  • A549 cells
  • Cyclooxygenase-2
  • IKKα/β
  • NF-κB
  • p44/42 MAPK
  • Raf-1
  • Ras
  • YC-1

ASJC Scopus subject areas

  • Pharmacology

Cite this

@article{12488eb9b74a4ac4abad3a6d37089418,
title = "Involvement of Ras/Raf-1/p44/42 MAPK in YC-1-induced cyclooxygenase-2 expression in human pulmonary epithelial cells",
abstract = "Our previous study demonstrated that 3-(5′-hydroxymethyl-2′-furyl)-1-benzylindazole (YC-1) might activate the soluble guanylate cyclase (sGC)/cGMP/protein kinase G (PKG) pathway to induce cyclooxygenase-2 (COX-2) expression in human pulmonary epithelial cells (A549). In this study, we further investigated the role of Raf-1 in YC-1-induced nuclear factor-κB (NF-κB) activation and COX-2 expression in A549 cells. YC-1-induced COX-2 expression was attenuated by a Raf-1 inhibitor (GW 5074) in a concentration-dependent manner. Treatment of A549 cells with YC-1 or 8-bromo-cGMP, a cell-permeable cGMP analogue, induced Raf-1 Ser338 phosphorylation in a time-dependent manner. YC-1-mediated Raf-1 activation was inhibited by an sGC inhibitor (ODQ), a PKG inhibitor (KT-5823), a Ras inhibitor (manumycin A), a dominant negative Ras mutant (RasN17), a protein kinase C-α (PKC-α) inhibitor (Ro 32-0432), and a phosphoinositide-3-OH-kinase (PI3K) inhibitor (LY 294002). Pretreatment of A549 cells with either manumycin A or GW 5074 attenuated YC-1-induced p44/42 MAPK activation. The YC-1-mediated increase in IKKα/β activation and κB-luciferase activity were attenuated by GW 5074, a MAPK/ERK kinase (MEK) inhibitor (PD 98059), and an ERK2 inhibitor (AG 126). Furthermore, YC-1-induced COX-2 promoter activity was also inhibited by GW 5074, PD 98059, and AG 126. These results indicate that YC-1 might activate the sGC/cGMP/PKG pathway to elicit Ras/Raf-1/p44/42 MAPK activation, which in turn induces IKKα/β and NF-κB activation, and ultimately causes COX-2 expression in A549 cells. Moreover, PKC-α and PI3K signal might be involved in YC-1-induced Raf-1 activation.",
keywords = "A549 cells, Cyclooxygenase-2, IKKα/β, NF-κB, p44/42 MAPK, Raf-1, Ras, YC-1",
author = "Chang, {Ming Shyan} and Chen, {Bing Chang} and Weng, {Chih Ming} and Lee, {Wen Sen} and Lin, {Chien Huang}",
year = "2009",
month = "10",
doi = "10.1016/j.phrs.2009.03.015",
language = "English",
volume = "60",
pages = "247--253",
journal = "Pharmacological Research",
issn = "1043-6618",
publisher = "Academic Press Inc.",
number = "4",

}

TY - JOUR

T1 - Involvement of Ras/Raf-1/p44/42 MAPK in YC-1-induced cyclooxygenase-2 expression in human pulmonary epithelial cells

AU - Chang, Ming Shyan

AU - Chen, Bing Chang

AU - Weng, Chih Ming

AU - Lee, Wen Sen

AU - Lin, Chien Huang

PY - 2009/10

Y1 - 2009/10

N2 - Our previous study demonstrated that 3-(5′-hydroxymethyl-2′-furyl)-1-benzylindazole (YC-1) might activate the soluble guanylate cyclase (sGC)/cGMP/protein kinase G (PKG) pathway to induce cyclooxygenase-2 (COX-2) expression in human pulmonary epithelial cells (A549). In this study, we further investigated the role of Raf-1 in YC-1-induced nuclear factor-κB (NF-κB) activation and COX-2 expression in A549 cells. YC-1-induced COX-2 expression was attenuated by a Raf-1 inhibitor (GW 5074) in a concentration-dependent manner. Treatment of A549 cells with YC-1 or 8-bromo-cGMP, a cell-permeable cGMP analogue, induced Raf-1 Ser338 phosphorylation in a time-dependent manner. YC-1-mediated Raf-1 activation was inhibited by an sGC inhibitor (ODQ), a PKG inhibitor (KT-5823), a Ras inhibitor (manumycin A), a dominant negative Ras mutant (RasN17), a protein kinase C-α (PKC-α) inhibitor (Ro 32-0432), and a phosphoinositide-3-OH-kinase (PI3K) inhibitor (LY 294002). Pretreatment of A549 cells with either manumycin A or GW 5074 attenuated YC-1-induced p44/42 MAPK activation. The YC-1-mediated increase in IKKα/β activation and κB-luciferase activity were attenuated by GW 5074, a MAPK/ERK kinase (MEK) inhibitor (PD 98059), and an ERK2 inhibitor (AG 126). Furthermore, YC-1-induced COX-2 promoter activity was also inhibited by GW 5074, PD 98059, and AG 126. These results indicate that YC-1 might activate the sGC/cGMP/PKG pathway to elicit Ras/Raf-1/p44/42 MAPK activation, which in turn induces IKKα/β and NF-κB activation, and ultimately causes COX-2 expression in A549 cells. Moreover, PKC-α and PI3K signal might be involved in YC-1-induced Raf-1 activation.

AB - Our previous study demonstrated that 3-(5′-hydroxymethyl-2′-furyl)-1-benzylindazole (YC-1) might activate the soluble guanylate cyclase (sGC)/cGMP/protein kinase G (PKG) pathway to induce cyclooxygenase-2 (COX-2) expression in human pulmonary epithelial cells (A549). In this study, we further investigated the role of Raf-1 in YC-1-induced nuclear factor-κB (NF-κB) activation and COX-2 expression in A549 cells. YC-1-induced COX-2 expression was attenuated by a Raf-1 inhibitor (GW 5074) in a concentration-dependent manner. Treatment of A549 cells with YC-1 or 8-bromo-cGMP, a cell-permeable cGMP analogue, induced Raf-1 Ser338 phosphorylation in a time-dependent manner. YC-1-mediated Raf-1 activation was inhibited by an sGC inhibitor (ODQ), a PKG inhibitor (KT-5823), a Ras inhibitor (manumycin A), a dominant negative Ras mutant (RasN17), a protein kinase C-α (PKC-α) inhibitor (Ro 32-0432), and a phosphoinositide-3-OH-kinase (PI3K) inhibitor (LY 294002). Pretreatment of A549 cells with either manumycin A or GW 5074 attenuated YC-1-induced p44/42 MAPK activation. The YC-1-mediated increase in IKKα/β activation and κB-luciferase activity were attenuated by GW 5074, a MAPK/ERK kinase (MEK) inhibitor (PD 98059), and an ERK2 inhibitor (AG 126). Furthermore, YC-1-induced COX-2 promoter activity was also inhibited by GW 5074, PD 98059, and AG 126. These results indicate that YC-1 might activate the sGC/cGMP/PKG pathway to elicit Ras/Raf-1/p44/42 MAPK activation, which in turn induces IKKα/β and NF-κB activation, and ultimately causes COX-2 expression in A549 cells. Moreover, PKC-α and PI3K signal might be involved in YC-1-induced Raf-1 activation.

KW - A549 cells

KW - Cyclooxygenase-2

KW - IKKα/β

KW - NF-κB

KW - p44/42 MAPK

KW - Raf-1

KW - Ras

KW - YC-1

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U2 - 10.1016/j.phrs.2009.03.015

DO - 10.1016/j.phrs.2009.03.015

M3 - Article

C2 - 19717011

AN - SCOPUS:69249202636

VL - 60

SP - 247

EP - 253

JO - Pharmacological Research

JF - Pharmacological Research

SN - 1043-6618

IS - 4

ER -