Effects of the Helicobacter pylori Virulence Factor CagA and Ammonium Ion on Mucins in AGS Cells
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Purpose: To investigate the effects of Helicobacter pylori (H. pylori)-CagA and the urease metabolite NH4 + on mucin expression inAGS cells. Materials and Methods: AGS cells were transfected with CagA and/or treated with different concentrations of NH4CL. Mucingene and protein expression was assessed by qPCR and immunofluorescence assays, respectively. Results: CagA significantly upregulated MUC5AC, MUC2, and MUC5B expression in AGS cells, but did not affect E-cadherin andMUC6 expression. MUC5AC, MUC6, and MUC2 expression in AGS cells increased with increasing NH4 + concentrations untilreaching a peak level at 15 mM. MUC5B mRNA expression in AGS cells (NH4 + concentration of 15 mM) was significantly higherthan that at 0, 5, and 10 mM NH4 + . No changes in E-cadherin expression in AGS cells treated with NH4 + were noted, except at 20mM. The expression of MUC5AC, MUC2, and MUC6 mRNA in CagA-transfected AGS cells at an NH4 + concentration of 15 mM wassignificantly higher than that at 0 mM, and decreased at higher concentrations. The expression of MUC5B mRNA increased withincreases in NH4 + concentration, and was significantly higher compared to that in untreated cells. No significant change in the expressionof E-cadherin mRNA in CagA-transfected AGS cells was observed. Immunofluorescence assays confirmed the observedchanges. Conclusion: H. pylori may affect the expression of MUC5AC, MUC2, MUC5B, and MUC6 in AGS cells via CagA and/or NH4 + , butnot E-cadherin.
Abstract
Purpose: To investigate the effects of Helicobacter pylori (H. pylori)-CagA and the urease metabolite NH4 + on mucin expression inAGS cells. Materials and Methods: AGS cells were transfected with CagA and/or treated with different concentrations of NH4CL. Mucingene and protein expression was assessed by qPCR and immunofluorescence assays, respectively. Results: CagA significantly upregulated MUC5AC, MUC2, and MUC5B expression in AGS cells, but did not affect E-cadherin andMUC6 expression. MUC5AC, MUC6, and MUC2 expression in AGS cells increased with increasing NH4 + concentrations untilreaching a peak level at 15 mM. MUC5B mRNA expression in AGS cells (NH4 + concentration of 15 mM) was significantly higherthan that at 0, 5, and 10 mM NH4 + . No changes in E-cadherin expression in AGS cells treated with NH4 + were noted, except at 20mM. The expression of MUC5AC, MUC2, and MUC6 mRNA in CagA-transfected AGS cells at an NH4 + concentration of 15 mM wassignificantly higher than that at 0 mM, and decreased at higher concentrations. The expression of MUC5B mRNA increased withincreases in NH4 + concentration, and was significantly higher compared to that in untreated cells. No significant change in the expressionof E-cadherin mRNA in CagA-transfected AGS cells was observed. Immunofluorescence assays confirmed the observedchanges. Conclusion: H. pylori may affect the expression of MUC5AC, MUC2, MUC5B, and MUC6 in AGS cells via CagA and/or NH4 + , butnot E-cadherin.
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