Limits...
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.

Show MeSH

Related in: MedlinePlus

Effect of inhibition of tyrosine kinase and MEK activity on IL-1β-stimulated gastric epithelial cell proliferation. AGS cells were treated with 10 ng/ml IL-1β for 24 hours in the presence of the tyrosine kinase inhibitor genistein (100 μM) or the MEK inhibitor PD 98059 (25 μM). Proliferation was assessed by [3H]thymidine incorporation. Studies were performed in the presence of anti-GM-CSF antibody (5 μg/ml). Results expressed as means ± standard deviation. *P < 0.01 vs control
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC103665&req=5

Figure 5: Effect of inhibition of tyrosine kinase and MEK activity on IL-1β-stimulated gastric epithelial cell proliferation. AGS cells were treated with 10 ng/ml IL-1β for 24 hours in the presence of the tyrosine kinase inhibitor genistein (100 μM) or the MEK inhibitor PD 98059 (25 μM). Proliferation was assessed by [3H]thymidine incorporation. Studies were performed in the presence of anti-GM-CSF antibody (5 μg/ml). Results expressed as means ± standard deviation. *P < 0.01 vs control

Mentions: The specific inhibitors genistein, which inhibits tyrosine kinases and PD 98059, which inhibits MAP kinase kinase (MEK), and thus inhibits the ERK-pathway, were used to assess the possible intracellular pathways mediating the effects of IL-1β. In order to examine the effects of IL-1β distinct from those of GM-CSF, these experiments were performed in the presence of the anti-GM-CSF neutralising antibody. As shown in figure 5, neither genistein nor PD 98059 altered unstimulated [3H]thymidine incorporation. Genistein completely abolished the IL-1β-stimulation of proliferation. Inhibition of MEK with PD 98059 reduced IL-1β-stimulated proliferation by 58 ± 5% (P < 0.01) (figure 5) but did not completely abolish the growth stimulatory action of IL-1β. Further increases to supra-maximal concentrations of PD 98050 did not further inhibit IL-1β-stimulated proliferation (data not shown).


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

Beales IL - BMC Gastroenterol (2002)

Effect of inhibition of tyrosine kinase and MEK activity on IL-1β-stimulated gastric epithelial cell proliferation. AGS cells were treated with 10 ng/ml IL-1β for 24 hours in the presence of the tyrosine kinase inhibitor genistein (100 μM) or the MEK inhibitor PD 98059 (25 μM). Proliferation was assessed by [3H]thymidine incorporation. Studies were performed in the presence of anti-GM-CSF antibody (5 μg/ml). Results expressed as means ± standard deviation. *P < 0.01 vs control
© Copyright Policy
Related In: Results  -  Collection

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

Figure 5: Effect of inhibition of tyrosine kinase and MEK activity on IL-1β-stimulated gastric epithelial cell proliferation. AGS cells were treated with 10 ng/ml IL-1β for 24 hours in the presence of the tyrosine kinase inhibitor genistein (100 μM) or the MEK inhibitor PD 98059 (25 μM). Proliferation was assessed by [3H]thymidine incorporation. Studies were performed in the presence of anti-GM-CSF antibody (5 μg/ml). Results expressed as means ± standard deviation. *P < 0.01 vs control
Mentions: The specific inhibitors genistein, which inhibits tyrosine kinases and PD 98059, which inhibits MAP kinase kinase (MEK), and thus inhibits the ERK-pathway, were used to assess the possible intracellular pathways mediating the effects of IL-1β. In order to examine the effects of IL-1β distinct from those of GM-CSF, these experiments were performed in the presence of the anti-GM-CSF neutralising antibody. As shown in figure 5, neither genistein nor PD 98059 altered unstimulated [3H]thymidine incorporation. Genistein completely abolished the IL-1β-stimulation of proliferation. Inhibition of MEK with PD 98059 reduced IL-1β-stimulated proliferation by 58 ± 5% (P < 0.01) (figure 5) but did not completely abolish the growth stimulatory action of IL-1β. Further increases to supra-maximal concentrations of PD 98050 did not further inhibit IL-1β-stimulated proliferation (data not shown).

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