cDNA array analysis of cag pathogenicity island-associated Helicobacter pylori epithelial cell response genes.
Helicobacter pylori strains containing the cag pathogenicity island (PAI) induce NF-kappaB activation and interleukin-8 secretion in gastric epithelial cells. The aim of this study was to investigate changes in epithelial gene expression induced by cag PAI-positive and -negative strains of H. pylori using high-density cDNA array hybridization technology. Radio-labeled cDNA prepared from H. pylori-infected Kato 3 gastric epithelial cells was hybridized to high-density cDNA arrays to identify changes in epithelial gene expression compared to noninfected controls. In vivo expression of selected, differentially expressed genes was examined by reverse transcription-PCR analysis of H. pylori-positive and -negative gastric mucosa. Screening of ca. 57,800 cDNAs identified 208 known genes and 48 novel genes and/or expressed sequence tags of unknown function to be differentially expressed in Kato 3 cells following H. pylori infection. Marked differences in gene expression profiles were observed following cag PAI-positive and cag PAI-negative infection with 15 novel cDNAs and 92 known genes being differentially expressed. H. pylori was found to change the expression of genes encoding growth factors and cytokine/chemokines and their receptors, apoptosis proteins, transcription factors and metalloprotease-disintegrin proteins (ADAMs), and tissue inhibitors of metalloproteinases. Gastric differential expression of selected known genes (amphiregulin and ADAM 10) and a novel gene (HPYR1) was confirmed in vivo in patients with H. pylori infection. Confirmation of the in vivo expression of selected genes demonstrates the usefulness of this approach for investigating pathogen-induced changes in host gene expression.
Cox
J M
JM
Molecular Medicine Unit, St. James's University Hospital, Leeds LS9 7TF, United Kingdom.
Clayton
C L
CL
Tomita
T
T
Wallace
D M
DM
Robinson
P A
PA
Crabtree
J E
JE
D051722:ADAM Proteins; D000072197:ADAM10 Protein; D066258:Amphiregulin; D053829:Amyloid Precursor Protein Secretases; D002460:Cell Line; D004269:DNA, Bacterial; D018076:DNA, Complementary; D066255:EGF Family of Proteins; D004847:Epithelial Cells / Q000166:cytology; D005753:Gastric Mucosa / Q000166:cytology / Q000382:microbiology / Q000473:pathology; D015870:Gene Expression; D020869:Gene Expression Profiling; D005798:Genes, Bacterial / Q000502:physiology*; D006023:Glycoproteins / Q000235:genetics; D006133:Growth Substances / Q000235:genetics; D016481:Helicobacter Infections / Q000235:genetics* / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D016480:Helicobacter pylori / Q000235:genetics* / Q000472:pathogenicity; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007700:Kinetics; D008565:Membrane Proteins / Q000235:genetics; D008666:Metalloendopeptidases / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods
C587848:AREG protein, human; D066258:Amphiregulin; D004269:DNA, Bacterial; D018076:DNA, Complementary; D066255:EGF Family of Proteins; D006023:Glycoproteins; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human