title: Extracellular signal-regulated protein kinases mediate H(+),K(+)-ATPase alpha-subunit gene expression.
abstract: Extracellular signal-regulated protein kinases (ERKs) are important in many cellular functions. We and others have previously reported that prolonged exposure of gastric parietal cells to epidermal growth factor (EGF) enhanced gastric acid secretion stimulated by secretagogues via ERKs. In this study, we examined whether ERKs regulated H(+),K(+)-ATPase alpha-subunit gene expression using a gastric cancer cell line, AGS. EGF induced ERK activity time- and dose-dependently with a maximal effect observed at 10 min and 10 nM, respectively. The MEK inhibitors, U0126 and PD-98059, dose-dependently inhibited the ERK activity stimulated by EGF. To test H(+),K(+)-ATPase alpha-subunit gene expression, we transfected AGS cells with a plasmid containing a canine H(+),K(+)-ATPase alpha-subunit gene promoter fused to a luciferase reporter gene. EGF induced luciferase activity in transfected cells; this effect was inhibited by the MEK inhibitors, suggesting that EGF-induced gene expression involved the ERK pathway. When AGS cells were transfected with the reporter plasmids in conjunction with an expression vector encoding constitutively active MEK1, luciferase activity was strongly enhanced; this effect was attenuated by the MEK inhibitors or by an additional cotransfection of dominant negative MEK1. Taken together, our results led us to conclude that the ERK pathway may mediate H(+),K(+)-ATPase alpha-subunit gene expression, contributing to gastric acid secretion in parietal cells.
authors.affiliation: Second Department of Internal Medicine, Showa University School of Medicine, 1-5-8, Hatanodai, Shinagawa-ku, Tokyo, 142-8666, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kusayanagi
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Takeuchi
authors.forename: Andrea
authors.initials: A
authors.lastname: Todisco
authors.forename: Keiji
authors.initials: K
authors.lastname: Mitamura
mesh_terms: D002070:Butadienes / Q000494:pharmacology; D002460:Cell Line; D005419:Flavonoids / Q000494:pharmacology; D005744:Gastric Acid / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D017506:H(+)-K(+)-Exchanging ATPase / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008156:Luciferases / Q000235:genetics; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D009570:Nitriles / Q000494:pharmacology; D010295:Parietal Cells, Gastric / Q000201:enzymology / Q000378:metabolism; D011401:Promoter Regions, Genetic; D021122:Protein Subunits; D015398:Signal Transduction; D014162:Transfection
chemical_list: D002070:Butadienes; D005419:Flavonoids; D009570:Nitriles; D021122:Protein Subunits; C113580:U 0126; D008156:Luciferases; D020928:Mitogen-Activated Protein Kinases; D017506:H(+)-K(+)-Exchanging ATPase; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one