Yonsei Med J.  2011 May;52(3):522-526. 10.3349/ymj.2011.52.3.522.

Effect of Pertussis Toxin and Herbimycin A on Proteinase-Activated Receptor 2-Mediated Cyclooxygenase 2 Expression in Helicobacter pylori-Infected Gastric Epithelial AGS Cells

• Seo JH
• Seo JY
• Chung HY
• Kim H

• KMID: 1777026
• DOI: http://doi.org/10.3349/ymj.2011.52.3.522

Abstract

Helicobacter pylori (H. pylori) is an important risk factor for chronic gastritis, peptic ulcer, and gastric cancer. Proteinase-activated receptor 2 (PAR2), subgroup of G-protein coupled receptor family, is highly expressed in gastric cancer, and chronic expression of cyclooxygenase-2 (COX-2) plays an important role in H. pylori-associated gastric carcinogenesis and inflammation. We previously demonstrated that H. pylori induced the expression of PAR2 and COX-2 in gastric epithelial cells. Present study aims to investigate whether COX-2 expression induced by H. pylori in Korean isolates is mediated by PAR2 via activation of Gi protein and Src kinase in gastric epithelial AGS cells. Results showed that H. pylori-induced COX-2 expression was inhibited in the cells transfected with antisense oligonucleotide for PAR2 or treated with Gi protein blocker pertussis toxin, Src kinase inhibitor herbimycin A and soybean trypsin inbitor, indicating that COX-2 expression is mediated by PAR2 through activation of Gi protein and Src kinase in gastric epithelial cells infected with H. pylori in Korean isolates. Thus, targeting the activation of PAR2 may be beneficial for prevention or treatment of gastric inflammation and carcinogenesis associated with H. pylori infection.

Keyword

Helicobacter pylori; cyclooxygenase-2; proteinase-activated receptor 2; AGS cells

MeSH Terms

Benzoquinones/*pharmacology
Cell Line, Tumor
Cyclooxygenase 2/genetics/*metabolism
Epithelial Cells/enzymology/metabolism/microbiology
GTP-Binding Protein alpha Subunits, Gi-Go/metabolism
Gastric Mucosa/enzymology/metabolism/*microbiology
*Helicobacter pylori
Humans
Lactams, Macrocyclic/*pharmacology
Oligonucleotides, Antisense
Pertussis Toxin/*pharmacology
RNA, Messenger/metabolism
Receptor, PAR-2/*physiology
src-Family Kinases/metabolism

Figure

• 

  Fig. 1 H. pylori-induced expressions of PAR2 and COX-2 are inhibited in AGS cells transfected with PAR2 AS ODN. AGS cells were seeded in 6-well culture plates at 5×105 cells per well transfected with S or AS ODNs for PAR-2 for 16 h. The bacterial cells were added to the cultured cells at a bacterium/cell ratio of 100 : 1 for 12 h (for mRNA) or 24 h (for protein). The expressions of mRNA and protein for PAR2 (A) and COX-2 (B) were determined by RT-PCR and Western blotting, respectively. PAR2, proteinase-activated receptor 2; COX-2, cyclooxygenase-2.

• 

  Fig. 2 H. pylori-induced expression of COX-2 is inhibited in AGS cells treated with soybean trypsin inhibitor (SBTI), pertussis toxin and herbimycin A. AGS cells in 6-well culture plates were treated with SBTI (1, 2, 5 nM) (A), or pertussis toxin (400 ng/mL) or herbimycin A (10 µM) (B) for 1 h and cultured in the presence of H. pylori at a bacterium/cell ratio of 100 : 1 for 12 h (for mRNA) or 24 h (for protein). COX-2 mRNA expression levels were determined by RT-PCR and at protein level by Western blotting. COX-2, cyclooxygenase-2.

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