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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 cytokine inhibition on IL-1β-stimulation of [3H]thymidine into gastric epithelial cells AGS cells were treated with 10 ng/ml IL-1β for 24 hours plus either interleukin-1 receptor antagonist (IL-1RA 500 ng/ml), neutralising anti-IL-8 antibody (10 μg/ml) or neutralising anti-GM-CSF antibody (5 μg/ml). DNA synthesis was assessed by [3H]thymidine incorporation. Results expressed as means ± standard deviation. *P < 0.01 vs. IL-1β stimulation in absence of inhibitor.
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Figure 3: Effect of cytokine inhibition on IL-1β-stimulation of [3H]thymidine into gastric epithelial cells AGS cells were treated with 10 ng/ml IL-1β for 24 hours plus either interleukin-1 receptor antagonist (IL-1RA 500 ng/ml), neutralising anti-IL-8 antibody (10 μg/ml) or neutralising anti-GM-CSF antibody (5 μg/ml). DNA synthesis was assessed by [3H]thymidine incorporation. Results expressed as means ± standard deviation. *P < 0.01 vs. IL-1β stimulation in absence of inhibitor.

Mentions: Pretreatment of the cells with interleukin-1 receptor antagonist abolished the stimulatory effects of IL-1β on [3H]thymidine incorporation (Figure 3). Previous studies have shown that IL-1β can activate gastric epithelial cells, including AGS cells, to secrete other cytokines, particularly interleukin-8 and granulocyte-macrophage colony stimulating factor (GM-CSF)[31,32]. Therefore further studies were undertaken to assess if the stimulatory actions of IL-1β were mediated by either of these two cytokines. Neutralising antibodies to either IL-8 or GM-CSF had no effect on unstimulated [3H]thymidine incorporation. Neutralisation of IL-8 had no effect on IL-1β-stimuated growth but the anti-GM-CSF antibody reduced IL-1β-stimulated proliferation by 31 ± 4 % (P < 0.01) (figure 3).


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

Beales IL - BMC Gastroenterol (2002)

Effect of cytokine inhibition on IL-1β-stimulation of [3H]thymidine into gastric epithelial cells AGS cells were treated with 10 ng/ml IL-1β for 24 hours plus either interleukin-1 receptor antagonist (IL-1RA 500 ng/ml), neutralising anti-IL-8 antibody (10 μg/ml) or neutralising anti-GM-CSF antibody (5 μg/ml). DNA synthesis was assessed by [3H]thymidine incorporation. Results expressed as means ± standard deviation. *P < 0.01 vs. IL-1β stimulation in absence of inhibitor.
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Related In: Results  -  Collection

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

Figure 3: Effect of cytokine inhibition on IL-1β-stimulation of [3H]thymidine into gastric epithelial cells AGS cells were treated with 10 ng/ml IL-1β for 24 hours plus either interleukin-1 receptor antagonist (IL-1RA 500 ng/ml), neutralising anti-IL-8 antibody (10 μg/ml) or neutralising anti-GM-CSF antibody (5 μg/ml). DNA synthesis was assessed by [3H]thymidine incorporation. Results expressed as means ± standard deviation. *P < 0.01 vs. IL-1β stimulation in absence of inhibitor.
Mentions: Pretreatment of the cells with interleukin-1 receptor antagonist abolished the stimulatory effects of IL-1β on [3H]thymidine incorporation (Figure 3). Previous studies have shown that IL-1β can activate gastric epithelial cells, including AGS cells, to secrete other cytokines, particularly interleukin-8 and granulocyte-macrophage colony stimulating factor (GM-CSF)[31,32]. Therefore further studies were undertaken to assess if the stimulatory actions of IL-1β were mediated by either of these two cytokines. Neutralising antibodies to either IL-8 or GM-CSF had no effect on unstimulated [3H]thymidine incorporation. Neutralisation of IL-8 had no effect on IL-1β-stimuated growth but the anti-GM-CSF antibody reduced IL-1β-stimulated proliferation by 31 ± 4 % (P < 0.01) (figure 3).

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