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01-04-2014 | Original Article

Helicobacter pylori urease and flagellin alter mucin gene expression in human gastric cancer cells

Authors: Michaël Perrais, Christel Rousseaux, Marie-Paule Ducourouble, René Courcol, Pascal Vincent, Nicolas Jonckheere, Isabelle Van Seuningen

Published in: Gastric Cancer | Issue 2/2014

Abstract

Background

Helicobacter pylori (Hp), which is one of the causative agents in human gastric adenocarcinoma, is known to interact with mucous gel and alter mucin gene expression. The aim of this work was to study, using an in vitro model of cell infection, the effects of urease, flagellin, and CagA virulence factors on the regulation of the four 11p15 mucin genes (MUC2, MUC5AC, MUC5B, and MUC6).

Methods

KATO-III and AGS gastric cancer cells were infected for 1, 3 or 6 h with Hp wild-type strains (ATCC 43504, N6, and SS1) or corresponding isogenic mutants deficient for urease subunit B, flagellin subunit A, and CagA. mRNA levels of MUC2, MUC5B, MUC5AC and MUC6 were assessed by RT-PCR, and functional activity of their promoters was measured by transient transfection assays.

Results

Infection of KATO-III cells with Hp wild-type strains resulted in an early (at 1 h) transient expression of MUC2, MUC5AC, and MUC6 mRNA concomitant with those of interleukin (IL)-1β, IL-8, and TNF-α cytokines. In these cells, the UreB⁻ isogenic mutant induced strong activation of MUC5AC expression, and UreB-responsive elements were located in the −486/−1 region of the promoter. FlaA⁻ and CagA⁻ mutants had no effect on mucin gene mRNA levels in KATO-III cells. In AGS cells, Hp-responsive elements were identified in all promoters, and overexpression of NF-κB induced upregulation of MUC5AC promoter activity when infected with the UreB⁻ isogenic mutant.

Conclusion

These results indicate that Hp infection of gastric cancer cells alters 11p15 mucin gene transcription and that MUC5AC downregulation is mediated by urease virulence factor.

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Title: Helicobacter pylori urease and flagellin alter mucin gene expression in human gastric cancer cells
Authors: Michaël Perrais
Christel Rousseaux
Marie-Paule Ducourouble
René Courcol
Pascal Vincent
Nicolas Jonckheere
Isabelle Van Seuningen
Publication date: 01-04-2014
Publisher: Springer Japan
Published in: Gastric Cancer / Issue 2/2014
Print ISSN: 1436-3291
Electronic ISSN: 1436-3305
DOI: https://doi.org/10.1007/s10120-013-0267-5

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