Helicobacter pylori potentiates epithelial:mesenchymal transition in gastric cancer: links to soluble HB-EGF, gastrin and matrix metalloproteinase-7

Abstract

Background and aimsHelicobacter pylori (H pylori) infection is a major risk factor in the development of distal gastric adenocarcinoma. Development of the invasive phenotype is associated with the phenomenon of epithelial:mesenchymal transition (EMT). Soluble heparin-binding epidermal growth factor (HB-EGF) has been implicated in this process. A study was undertaken to investigate the possibility that matrix metalloproteinase (MMP)-7 is upregulated in H pylori infection as a result of hypergastrinaemia, which may enhance shedding of HB-EGF and contribute towards EMT in gastric adenocarcinoma cell lines.MethodsThree gastric epithelial cell lines (AGS, MGLVA1 and ST16) were co-cultured with the pathogenic H pylori strain 60190 and non-pathogenic strain Tx30a in an in vitro infection model. Gene expression was quantified by real-time PCR, HB-EGF shedding by ELISA and protein expression by immunofluorescence or immunohistochemistry. The INS-GAS mouse, a transgenic mouse model of gastric carcinogenesis which overexpresses amidated gastrin, was used to investigate the in vivo relationship between HB-EGF, MMP-7, gastrin and EMT.ResultsThe pathogenic strain of H pylori significantly upregulated EMT-associated genes Snail, Slug and vimentin in all three gastric cell lines to a greater degree than the non-pathogenic strain. Pathogenic H pylori also upregulated HB-EGF shedding, a factor implicated in EMT, which was partially dependent on both gastrin and MMP-7 expression. Gastrin and MMP-7 siRNAs and MMP-7 neutralising antibody significantly reduced upregulation of HB-EGF shedding in H pylori infected gastric cell lines and reduced EMT gene expression. The effect of H pylori on EMT was also reversed by gastrin siRNA. Neutralisation of gastrin in the INS-GAS mouse model reduced expression of MMP-7, HB-EGF and key EMT proteins.ConclusionThe upregulation of MMP-7 by pathogenic H pylori is partially dependent on gastrin and may have a role in the development of gastric cancer, potentially through EMT, by indirectly increasing levels of soluble HB-EGF.

Zinc ions upregulate the hormone gastrin via an E-box motif in the proximal gastrin promoter

“…Infection of AGS and MGLVA1 cells by pathogenic Helicobacter pylori results in the upregulation of the EMT marker matrix metalloproteinase 7 (MMP7) and a consequent increase in soluble HB-EGF. Both outcomes are partially dependent on gastrin in vivo and in vitro (Yin et al 2010). Furthermore, gastrin has been shown to increase migration of AGS cells expressing CCK2R receptors via upregulation of MMP7 (Mishra et al 2010).…”

Lung Cancer-derived Galectin-1 Enhances Tumorigenic Potentiation of Tumor-associated Dendritic Cells by Expressing Heparin-binding EGF-like Growth Factor

“…HB-EGF, a member of the EGF family, has been described as a mitogenic factor of various types of cells, including cancer cells (9,26). Overexpression of HB-EGF is correlated with tumor progression and poor patient survival in ovarian, colon, head and neck squamous cell carcinoma, stomach, and breast cancers (27,28). In addition, up-regulation of HB-EGF has also been associated with chemoresistance to various chemotherapeutic drugs (29).…”

Gastric Cancer: Clinical Aspects, Epidemiology and Molecular Background

“…The upregulation of MMP7 is associated with lymph node invasion and shorter survival rates [61]. In cell lines, MMP7 is, together with gastrin, involved in the H. pylori-driven process of epithelial-mesenchymal transition (EMT) in the course of invasive GC development [62]. Neutralization of both gastrin and MMP7 prevented upregulation of heparin-binding epidermal growth factor that is a strong promotor of EMT.…”