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Transformed immortalized gastric epithelial cells by virulence factor CagA of Helicobacter pylori through Erk mitogen-activated protein kinase pathway

Oncogene volume 24, pages 3886–3895 (2005)Cite this article

Abstract

CagA of Helicobacter pylori is a protein that has been closely associated with gastric cancer and that can intervene with signal pathways in cells. Its precise relationship with the occurrence of gastric cancer, however, remains unclear. The purpose of this study is to investigate whether CagA can promote transformation of normal gastric epithelial cells and to consider via what mechanisms CagA may exert its effects. Transformed colonies were merged in soft-agarose medium after immortalized gastric epithelial cells were transfected with recombinant pLHCX retrovirus with cagA and/or dimethylhydrazine. The number of transformed colonies in the group containing cagA/pLHCX retrovirus, combined with a subthreshold dose of dimethylhydrazine, was more than that for cagA/pLHCX retrovirus or dimethylhydrazine at a subthreshold dose alone. For cagA-transfected cells, only IQGAP-2, R-Ras and B-Raf of the Ras/mitogen-activated protein kinase signal pathway were markedly increased, and the activity of extracellular signal-regulated kinase 1/2 (Erk1/2) kinase was significantly higher than that in dimethylhydrazine-transformed cells or control cells. However, no evidence of alteration of any other molecules of the Ras superfamily was observed in cagA-transfected cells. These findings suggest that CagA can transform gastric epithelial cells through activation of the Erk1/2 pathway; this mechanism may, however, be independent of Ras activation.

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Abbreviations

DMH:

1,2-dimethylhydrazine

SHP-2:

tyrosine phosphatase containing SH2 homolog

SV40 LT:

simian virus large T antigen

IVT:

in vitro transcription

PBS:

phosphate-buffered saline

TBS/T:

Tris buffer saline/Tween 20

PMSF:

phenylmethylsulfonyl fluoride

BSA:

bovine serum albumin

EDTA:

ethylenediamine tetraacetic acid

MAPK:

mitogen-activated protein kinase

Erk1/2:

extracellular signal-regulated kinase 1/2

SAPK:

stress-activated protein kinase

DMSO:

dimethyl sulfoxide

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Acknowledgements

We are very grateful to Mrs Jie Lin for her support. We thank Wenzhi Jiang for his technical assistance and the entire laboratory for fruitful discussions. This work was supported by the Natural Science Foundation of China (2001AA227111).

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Authors and Affiliations

  1. Gastroenterological Department, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China

    Yongliang Zhu & Qin Du

  2. Cancer Institute, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China

    Xian Zhong, Shu Zheng & Weizhen Xu

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Correspondence to Shu Zheng.

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Zhu, Y., Zhong, X., Zheng, S. et al. Transformed immortalized gastric epithelial cells by virulence factor CagA of Helicobacter pylori through Erk mitogen-activated protein kinase pathway. Oncogene 24, 3886–3895 (2005). https://doi.org/10.1038/sj.onc.1208551

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