Involvement of Cyclooxygenase-2–Prostaglandin E2 Pathway in Interleukin-8 Production in Gastric Cancer Cells

• DOI: 10.1007/s10620-006-9436-2
• Publication Date: 2006-10-31T16:33:26Z
• Authors: Hisashi Takehara; Jun-Ichi Iwamoto; Yuji Mizokami; Kimiko Takahashi; Toshiya Ootubo; Syuuhei Miura; Toshiaki Narasaka; Hiroki Takeyama; Takayuki Omata; Koichi Shimokobe; Masanori Ito; Takeshi Matsuoka
• Keywords: 0301 basic medicine; Sulfonamides; Arachidonic Acid; Cyclooxygenase 2 Inhibitors; Helicobacter pylori; Prostaglandin Antagonists; Xanthones; Biphenyl Compounds; Interleukin-8; Adenocarcinoma; Fibroblasts; Dinoprostone; 3. Good health; Gene Expression Regulation, Neoplastic; 03 medical and health sciences; Cyclooxygenase 2; Stomach Neoplasms; Cell Line, Tumor; Cyclic AMP; Humans; Receptors, Prostaglandin E; RNA, Messenger; Nitrobenzenes

Prostaglandin E(2) (PGE(2)) is thought to play an important role in both inflammatory and anti-inflammatory effects. The effect of PGE(2) on the proinflammatory chemokine interleukin-8 (IL-8) in the gastric epithelial cells has not been defined yet. A gastric cancer cell line (MKN45) and primary gastric fibroblasts were cocultured with Helicobacter pylori standard strain (NCTC11637). The expressions of IL-8 and cyclooxygenase 2 (COX-2) mRNA were examined by reverse transcription polymerase chain reaction (RT-PCR) amplification. The amount of IL-8 antigen secreted by the MKN45 cells and gastric fibroblasts was measured by enzyme-linked immunosorbent assay (ELISA). We examined the effects of H pylori stimulation on IL-8 and COX-2 expression levels and the effects of COX-2 inhibitor on H pylori-induced IL-8 production in the MKN45 cells and gastric fibroblasts. Furthermore, we examined the expressions of subtypes of PGE(2) receptors, the effects of arachidonic acid and PGE(2) on IL-8 production, and the effects of PGE(2) on the total cellular cyclic adenosine monophosphate (cAMP) in MKN45 cells. MKN45 cells and gastric fibroblasts expressed IL-8 and COX-2 mRNA under stimulation with H pylori. The MKN45 cells produced IL-8 and PGE(2) antigen into the culture medium with H pylori stimulation, and the production level of IL-8 and PGE(2) antigen decreased significantly with COX-2 inhibitor pretreatment (concentration: 50 muM). On the other hand, the gastric fibroblasts strongly produced IL-8 antigen even in the unstimulated condition, and the amount of IL-8 antigen was not affected by H pylori stimulation and/or COX-2 inhibitor pretreatment. The MKN45 cells expressed IL-8 mRNA and released IL-8 antigen slightly, and the expression level of IL-8 mRNA and the amount of IL-8 antigen increased significantly with PGE(2) treatment in a dose-dependent manner. PGE(2)-induced IL-8 production was inhibited by pretreatment with EP2 and EP4 antagonists. The MKN45 cells expressed EP2 and EP4 subtypes of PGE(2) receptors, and these expression levels were not affected by H pylori stimulation or PGE(2) treatment. The amount of IL-8 antigen increased slightly, but not significantly, with arachidonic acid treatment. PGE(2) treatment for 15 minutes increased the total cellular cAMP in the MKN45 cells. These results suggest that the COX-2-PGE(2) pathway may be involved in IL-8 production in gastric epithelial cells.