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. 2010 Jun 28;59(8):1037–1045. doi: 10.1136/gut.2009.199794

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© 2010, Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

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Effect of co-culture of gastric epithelial cells with Helicobacter pylori strain 60190 on matrix metalloproteinase 7 (MMP-7) expression. (a) MMP-7 gene expression following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains. *p<0.001 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (b) MMP-7 protein expression following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains. Cells were stained for MMP-7 (green fluorescence) and nuclei were counterstained with Hoechst (blue fluorescence). (i) AGS, (ii) AGS/H pylori 60190, (iii) AGS H pylori Tx30a. Magnification ×20. (c) MMP-7 protein expression level quantified by image analysis following co-culture of gastric epithelial cells with H pylori pathogenic (60190) and non-pathogenic (Tx30a) strains; *significantly higher MMP-7 protein expression level compared with untreated control cells (p<0.001). (d) MMP-7 and heparin-binding epidermal growth factor (HB-EGF) co-localisation: MMP-7 expression was stained by green fluorescence (i), HB-EGF by red fluorescence (ii), nuclei were stained with Hoechst blue fluorescence (iii). Merged image shows co-localisation of MMP-7 and HB-EGF (iv). Magnification ×20. (e) MMP-7 gene expression in MMP-7 siRNA treated gastric cell lines. MMP-7 siRNA significantly reduced MMP-7 mRNA expression in all three gastric cell lines tested. *p<0.001 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (f) HB-EGF gene expression in MMP-7 siRNA treated gastric cell lines exposed to H pylori 60190. MMP-7 siRNA significantly reduced HB-EGF shedding in all three gastric cell lines tested. *p<0.02, **p<0.03 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (g) HB-EGF gene expression in MMP-7 neutralising antibody treated gastric cell lines exposed to H pylori 60190. *p<0.03 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (h) HB-EGF shedding levels in gastric cell lines exposed to H pylori 60190 treated with an MMP-7 neutralising antibody. *p<0.04, **p<0.03 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence). (i) EMT gene expression in MMP-7 siRNA treated gastric cell lines exposed to H pylori strain 60190: Snail, p<0.01; Slug, p<0.005; vimentin, p<0.001 (n=3 replicates per condition and a representative graph is shown, error bars indicate 95% confidence).