Inhibition of human gastric H(+)-K(+)-ATPase alpha-subunit gene expression by Helicobacter pylori.
Clinical studies and in vitro data from isolated parietal cells suggest that acute Helicobacter pylori infection inhibits acid secretion. Gastric acidification is mediated by H(+)-K(+)-ATPase, an integral protein of parietal cell apical membranes. To test the hypothesis that H. pylori downregulates H(+)-K(+)-ATPase alpha-subunit (HKalpha) gene expression and to identify potential intracellular signaling pathways mediating such regulation, we transfected human gastric adenocarcinoma (AGS) cells with human and rat HKalpha 5'-flanking DNA fused to a luciferase reporter plasmid. Histamine caused dose-dependent, cimetidine-sensitive (10(-4) M) increases in cAMP, free intracellular Ca(2+), and HKalpha promoter activation in AGS cells. H. pylori infection of transfected AGS cells dose dependently inhibited basal and histamine-stimulated HKalpha promoter activity by 80% and 66%, respectively. H. pylori dose dependently inhibited phorbol myristate acetate-induced (10(-7) M) and staurosporine- (10(-7) M) and calphostin C-sensitive (5 x 10(-8) M) activation of HKalpha promoter. Also, H. pylori inhibited epidermal growth factor (EGF) (10(-8) M), genistein-sensitive (5 x 10(-5) M) activation of HKalpha promoter, reducing activity to 60% of basal level. These data suggest that H. pylori inhibits HKalpha gene expression via intracellular pathways involving protein kinases A and C and protein tyrosine kinase, AGS cells have functional histamine H(2) and EGF receptors, and transiently transfected AGS cells are a useful model for studying regulation of HKalpha gene expression.
Göõz
M
M
Department of Medicine, Medical University of South Carolina, Charleston, South Carolina 29425, USA.
Hammond
C E
CE
Larsen
K
K
Mukhin
Y V
YV
Smolka
A J
AJ
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002927:Cimetidine / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D015536:Down-Regulation; D015870:Gene Expression / Q000502:physiology*; D017506:H(+)-K(+)-Exchanging ATPase / Q000235:genetics*; D016481:Helicobacter Infections / Q000235:genetics; D016480:Helicobacter pylori / Q000502:physiology*; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000502:physiology; D051381:Rats; D013270:Stomach / Q000201:enzymology*; D014162:Transfection; D014407:Tumor Cells, Cultured
D007527:Isoenzymes; D002927:Cimetidine; D006632:Histamine; D000242:Cyclic AMP; D017506:H(+)-K(+)-Exchanging ATPase; D002118:Calcium