Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptional profiling of gastric epithelial cells infected with wild type or arginase-deficient Helicobacter pylori

ABSTRACT: We studied the differential gene expression in the gastric cell line AGS infected with wild type (WT) or arginase gene mutant (rocF) Helicobacter pylori (Hp). We found that the presence of the gene regulates the inflammatory responses in terms of IL8 and MIP-1B. We conclude that arginase may be involved in regulating inflammatory responses which improves the chances of survival against the human immune response. AGS cells were infected for 16 hours with WT, the rocF, or a complemented H. pylori strain (rocF+). The RNA was extracted with TRIzol, treated with DNAse I (both from invitrogen). cRNA was made with the Illumina TotalPrep RNA amplification Kit (ambion AMIL1791).

ORGANISM(S): Homo sapiens

SUBMITTER: Jovanny Zabaleta

PROVIDER: E-GEOD-39919 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Background: Helicobacter pylori is an important human pathogen that infects the human gastric mucosa leading to chronic inflammation which is responsible for severe gastroduodenal diseases. The Outer Inflammatory Protein A (OipA) of H. pylori is an important virulence factor. However, the role of OipA in gastric cancer and ulcer development is still not well-established. Phase variation within a CT dinucleotide repeat motif of oipA switches its expression “on” and “off”. This study aims to elucidate the role of OipA in H. pylori infection using oipA “on” and “off” clinical strains and oipA deletion mutant. Methods: Proteomics analysis was performed on AGS human gastric epithelial cell line post-infection with oipA “on” and “off” H. pylori using liquid chromatography / mass spectrometry (LC/MS). In addition, AGS apoptosis and cell cycle assays were performed. To confirm the findings, oipA deletion mutant (ΔoipA) was generated and expression of VacA was detected using Western blotting. Result: Expression of AGS proteins involve in human disease was suppressed / down-regulated post-infection with oipA “off” strains, as well as oipA mutant, compared to oipA “on” strains. Furthermore, oipA “off” strains and oipA mutant resulted in a higher level of apoptosis and G0/G1 cell-cycle arrest in AGS cells than oipA “on” strains. Interestingly, deletion of oipA was found to increase VacA production by the bacterium. Discussion: The capability of oipA “off” strains to induce apoptosis and suppress proteins having roles in human disease may suggest that these H. pylori may be less virulent or may even be protective against carcinogenesis compared to its oipA “on” counterparts. This may potentially explain the higher incidence of gastric cancer among East Asian which oipA “on” strains predominates. Conclusions: Data from this study strengthened our understanding of the role of H. pylori OipA as a virulence factor in chronic inflammation, metastasis and carcinogenesis.

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Transcriptional profiling of gastric epithelial cells infected with wild type or arginase-deficient Helicobacter pylori

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