Gastric Epithelial Cell Line AGS Research Articles

The Role of Nitric Oxide on Cigarette Smoke-induced Programmed Cell Death in the Gastric Mucosa

Background: This study was designed to investigate the effect of cigarette smoking on apoptosis in the gastric mucosa and the role of nitric oxide (NO) in the gas phase and extracts in the tar phase in this pathological process. Methods: Male Sprague-Dawley rats and human gastric epithelial cell line AGS were used in the study. Results: Cigarette smoking significantly increased apoptotic bodies in the rat gastric mucosa. However, neither filtered cigarette smoke, in which most of the substances in the tar phase were removed, nor oral administration of the two cigarette smoke extracts produced any effect on apoptosis. Interestingly, in this experiment pretreatment with a NO donor, the chloroform extract (CE) could significantly increase apoptosis. In in vitro study, only the CE induced DNA fragmentation, which could be elevated further by preincubation with a NO donor. The same extract also elevated inducible nitric oxide synthase (iNOS) activity. Inhibition of iNOS by NG-nitro-L-arginine methylester hydrochloride (L-NAME) partially abolished CE-induced apoptosis. Conclusions: These findings suggested that exogenous and endogenous NO had a synergistic effect with substances in the tar phase to induce programmed cell death in gastric epithelial cells both in vivo and in vitro.

Helicobacter pylori increases proliferation of gastric epithelial cells.

The direct and indirect effects of helicobacter pylori on cell kinetics of gastric epithelial cell line AGS were investigated by flow cytometric analysis of Ki-67 positive cells and by MTT assay. Flow cytometric analysis of Ki-67 positivity permits detection of cells that are in S-phase, whereas the MTT assay is a colometric measure of the number of viable cells. In the absence of added stimulants, 23.06 (4.88)% mean (SD) of AGS cells were Ki-67 positive. When cells were preincubated in the presence of H pylori, there was a significant increase in Ki-67 positivity (66.20 (7.89)%, p < 0.001). This increase was not seen in cells cultured in the presence of Campylobacter jejuni (24.63 (8.11)% or Escherichia coli (21.66 (9.78)%). Pre-incubation of AGS cells with supernatants from both H pylori and mitogen activated peripheral blood lymphocytes also increased the per cent of cells that were Ki-67 positive (72.93 (8.68) and 69.96 (12.35)%; p, 0.001) respectively. Similar results were also found in MTT assay. These data show that both H pylori directly and the immune/inflammatory response to H pylori indirectly can influence the rate of epithelial cell proliferation, suggesting this bacterium may be an initiating step in gastric carcinogenesis and an important co-carcinogenic factor in H pylori positive subjects.

Direct and indirect effects of H. pylori in cell cycle of gastric epithelial cell line AGS

Direct and indirect effects of H. pylori in cell cycle of gastric epithelial cell line AGS

Up-regulation of CD44 and ICAM-1 expression on gastric epithelial cells by H. pylori.

Interaction between lymphocytes and epithelial cells may play a key role in Helicobacter pylori (H. pylori)-associated gastric mucosal inflammation. This interaction process is at least partially mediated by various cell adhesion molecules. The aims of the present study were to assess using flow cytometric analysis whether H. pylori directly or supernatants from H. pylori-activated peripheral blood mononuclear cells (PBMC) can affect the expression of adhesion molecules on the gastric epithelial cell line AGS in vitro. The results showed that resting AGS cells expressed CD44 and ICAM-1. Co-culture of AGS with H. pylori or cytokine-rich supernatants from H. pylori-activated PBMC resulted in up-regulation of expression of CD44 and ICAM-1 on AGS cells. These data suggest that H. pylori directly and indirectly through inflammatory cytokines may contribute to alternations in adhesion molecule expression on gastric epithelial cells. This may be of pathological significance in H. pylori-associated gastric mucosal inflammation and carcinogenesis.