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Effect of interlukin-1beta on proliferation of gastric epithelial cells in culture.

Beales IL - BMC Gastroenterol (2002)

Bottom Line: Addition of neutralising antibody to GM-CSF reduced IL-1beta-stimulated proliferation by 31 +/- 4 %.GM-CSF alone significantly stimulated proliferation.Addition or neutralisation of IL-8 had no effect on basal or IL-1beta-stimulated proliferation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cell Biology School of Medicine, Health Policy and Practice, University of East Anglia Norwich, NR4 7TJ, United Kingdom. ibeales@uea.ac.uk

ABSTRACT

Background: Helicobacter pylori is the main risk factor for the development of non-cardia gastric cancer. Increased proliferation of the gastric mucosa is a feature of H. pylori infection. Mucosal interkeukin-1beta production is increased in H. pylori infection and IL-1beta genotypes associated with increased pro-inflammatory activity are risk factors for the development of gastric cancer. The effect of IL-1beta on gastric epithelial cell proliferation has been examined in this study.

Methods: AGS cells were cultured with IL-1beta. DNA synthesis was assed by [3H]thymidine incorporation and total viable cell numbers by MTT assay.

Results: IL-1beta dose dependently increased DNA synthesis and cell numbers. The enhanced proliferation was blocked by interleukin-1 receptor antagonist. Addition of neutralising antibody to GM-CSF reduced IL-1beta-stimulated proliferation by 31 +/- 4 %. GM-CSF alone significantly stimulated proliferation. Addition or neutralisation of IL-8 had no effect on basal or IL-1beta-stimulated proliferation. The tyrosine kinase inhibitor genistein completely blocked IL-1beta-stimulated proliferation and inhibition of the extracellular signal related kinase pathway with PD 98059 inhibited IL-1beta stimulated proliferation by 58 +/- 5 %.

Conclusions: IL-1beta stimulates proliferation in gastric epithelial cells. Autocrine stimulation by GM-CSF contributes to this proliferative response. Signalling via tyrosine kinase activity is essential to the mitogenic response to IL-1beta. The extracellular signal related kinase pathway is involved in, but not essential to downstream signalling. IL-1beta may contribute to the hyperproliferation seen in H. pylori- infected gastric mucosa, and be involved in the carcinogenic process.

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Effect of GM-CSF on proliferation of gastric epithelial cells. AGS cells were treated with increasing concentrations of GM-CSF for 24 hours. Cell proliferation was assessed by [3H]thymidine incorporation. Results expressed as means ± standard deviation. *P < 0.05, **P < 0.01 vs control
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Figure 4: Effect of GM-CSF on proliferation of gastric epithelial cells. AGS cells were treated with increasing concentrations of GM-CSF for 24 hours. Cell proliferation was assessed by [3H]thymidine incorporation. Results expressed as means ± standard deviation. *P < 0.05, **P < 0.01 vs control

Mentions: In view of the results obtained with the anti-GM-SCF antibody, the direct growth-stimulatory actions of GM-CSF were examined. GM-CSF had a potent growth stimulatory action on AGS cells: significant enhancement of [3H]thymidine incorporation was seen at all concentrations (0.001–100 ng/ml) of GM-CSF. GM-CSF itself appeared to be a more potent stimulant than IL-1β ; maximal stimulation of 108 ± 17 % above control was seen with 100 ng/ml of GM-CSF (Figure 4). The inhibitory action of the anti-GM-CSF antibody was confirmed by abolition of the growth stimulatory action of 1 ng/ml GM-CSF (data not shown). Previous studies have shown that IL-1β-stimulated GM-CSF release under similar conditions in AGS cells to be approximately 10–20 pg/well/24 hours [32]. To confirm the results obtained with the anti-IL-8 antibody, [3H]thymidine incorporation was measured in response to IL-8. No enhancement of proliferation was seen at any concentration of IL-8 (0.001–100 ng/ml) (data not shown). Under similar conditions IL-1β-stimulated IL-8 release is approximately 3000 pg/well/24 hours [31].


Effect of interlukin-1beta on proliferation of gastric epithelial cells in culture.

Beales IL - BMC Gastroenterol (2002)

Effect of GM-CSF on proliferation of gastric epithelial cells. AGS cells were treated with increasing concentrations of GM-CSF for 24 hours. Cell proliferation was assessed by [3H]thymidine incorporation. Results expressed as means ± standard deviation. *P < 0.05, **P < 0.01 vs control
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC103665&req=5

Figure 4: Effect of GM-CSF on proliferation of gastric epithelial cells. AGS cells were treated with increasing concentrations of GM-CSF for 24 hours. Cell proliferation was assessed by [3H]thymidine incorporation. Results expressed as means ± standard deviation. *P < 0.05, **P < 0.01 vs control
Mentions: In view of the results obtained with the anti-GM-SCF antibody, the direct growth-stimulatory actions of GM-CSF were examined. GM-CSF had a potent growth stimulatory action on AGS cells: significant enhancement of [3H]thymidine incorporation was seen at all concentrations (0.001–100 ng/ml) of GM-CSF. GM-CSF itself appeared to be a more potent stimulant than IL-1β ; maximal stimulation of 108 ± 17 % above control was seen with 100 ng/ml of GM-CSF (Figure 4). The inhibitory action of the anti-GM-CSF antibody was confirmed by abolition of the growth stimulatory action of 1 ng/ml GM-CSF (data not shown). Previous studies have shown that IL-1β-stimulated GM-CSF release under similar conditions in AGS cells to be approximately 10–20 pg/well/24 hours [32]. To confirm the results obtained with the anti-IL-8 antibody, [3H]thymidine incorporation was measured in response to IL-8. No enhancement of proliferation was seen at any concentration of IL-8 (0.001–100 ng/ml) (data not shown). Under similar conditions IL-1β-stimulated IL-8 release is approximately 3000 pg/well/24 hours [31].

Bottom Line: Addition of neutralising antibody to GM-CSF reduced IL-1beta-stimulated proliferation by 31 +/- 4 %.GM-CSF alone significantly stimulated proliferation.Addition or neutralisation of IL-8 had no effect on basal or IL-1beta-stimulated proliferation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cell Biology School of Medicine, Health Policy and Practice, University of East Anglia Norwich, NR4 7TJ, United Kingdom. ibeales@uea.ac.uk

ABSTRACT

Background: Helicobacter pylori is the main risk factor for the development of non-cardia gastric cancer. Increased proliferation of the gastric mucosa is a feature of H. pylori infection. Mucosal interkeukin-1beta production is increased in H. pylori infection and IL-1beta genotypes associated with increased pro-inflammatory activity are risk factors for the development of gastric cancer. The effect of IL-1beta on gastric epithelial cell proliferation has been examined in this study.

Methods: AGS cells were cultured with IL-1beta. DNA synthesis was assed by [3H]thymidine incorporation and total viable cell numbers by MTT assay.

Results: IL-1beta dose dependently increased DNA synthesis and cell numbers. The enhanced proliferation was blocked by interleukin-1 receptor antagonist. Addition of neutralising antibody to GM-CSF reduced IL-1beta-stimulated proliferation by 31 +/- 4 %. GM-CSF alone significantly stimulated proliferation. Addition or neutralisation of IL-8 had no effect on basal or IL-1beta-stimulated proliferation. The tyrosine kinase inhibitor genistein completely blocked IL-1beta-stimulated proliferation and inhibition of the extracellular signal related kinase pathway with PD 98059 inhibited IL-1beta stimulated proliferation by 58 +/- 5 %.

Conclusions: IL-1beta stimulates proliferation in gastric epithelial cells. Autocrine stimulation by GM-CSF contributes to this proliferative response. Signalling via tyrosine kinase activity is essential to the mitogenic response to IL-1beta. The extracellular signal related kinase pathway is involved in, but not essential to downstream signalling. IL-1beta may contribute to the hyperproliferation seen in H. pylori- infected gastric mucosa, and be involved in the carcinogenic process.

Show MeSH
Related in: MedlinePlus