East Asian-type Helicobacter pylori cytotoxin-associated gene A protein has a more significant effect on growth of rat gastric mucosal cells than the Western type

Hai Ying Fu; Kayoko Asahi; Yujiro Hayashi; Hiroshi Eguchi; Hiroaki Murata; Masahiko Tsujii; Shingo Tsuji; Takeshi Azuma; Sunao Kawano

doi:10.1111/j.1440-1746.2006.04531.x

East Asian-type Helicobacter pylori cytotoxin-associated gene A protein has a more significant effect on growth of rat gastric mucosal cells than the Western type

Hai Ying Fu, Kayoko Asahi, Yujiro Hayashi, Hiroshi Eguchi, Hiroaki Murata, Masahiko Tsujii, Shingo Tsuji, Takeshi Azuma, Sunao Kawano

Physiology & Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

Abstract

Background and Aim: Cytotoxin-associated gene A (CagA) protein from H. pylori was reported to be injected into host gastric epithelial cells via a bacterial type IV secretion system, thereby modifying signal transduction. It is classified into two major subtypes, Western and East Asian. The present study aimed to compare the effects of East Asian-type and Western-type CagA on host cell growth. Methods: A tetracycline (tet)-off system and cagA genes from Western and East Asian-type H. pylori (NCTC 11637 and F32) were transfected into untransformed rat gastric mucosal (RGM1) cells to establish RGM1-CagA cell lines in which CagA expression could be controlled by tetracycline. These cell lines were used to investigate the effect of CagA protein expression on cell growth with BrdU and water-soluble tetrazolium reagent (WST-8) assays. CagA expression, phosphorylation and extracellular signal-regulated kinase (ERK) activation were examined with immunoprecipitation and Western blotting analysis. Results: 5-Bromo-2′deoxyuridine (BrdU) and WST-8 assays demonstrated significant increases in DNA replication and RGM1 cell growth after CagA protein expression. ERK phosphorylation was enhanced when CagA protein was expressed in RGM1-CagA cells. Moreover, the East Asian-type CagA showed a significantly greater effect on ERK activation and host cell growth than the Western type. PD98059, a mitogen-activated protein kinase kinase (MEK) inhibitor, suppressed ERK phosphorylation and the CagA protein-induced increase in RGM1-CagA cell growth. Conclusions: CagA protein expression induces an increase in RGM1-CagA cell proliferation via the mitogen-activated protein kinase (MAPK) signal pathway. The East Asian-type CagA showed a significantly greater effect on cell growth than the Western type, suggesting that the East Asian CagA-positive strain may have an important role in pathogenesis.

Original language	English (US)
Pages (from-to)	355-362
Number of pages	8
Journal	Journal of Gastroenterology and Hepatology (Australia)
Volume	22
Issue number	3
DOIs	https://doi.org/10.1111/j.1440-1746.2006.04531.x
State	Published - Mar 2007

Keywords

CagA
Cell growth
Helicobacter pylori
MAPK
RGM1 cells

ASJC Scopus subject areas

Hepatology
Gastroenterology

Access to Document

10.1111/j.1440-1746.2006.04531.x

Cite this

Fu, H. Y., Asahi, K., Hayashi, Y., Eguchi, H., Murata, H., Tsujii, M., Tsuji, S., Azuma, T., & Kawano, S. (2007). East Asian-type Helicobacter pylori cytotoxin-associated gene A protein has a more significant effect on growth of rat gastric mucosal cells than the Western type. Journal of Gastroenterology and Hepatology (Australia), 22(3), 355-362. https://doi.org/10.1111/j.1440-1746.2006.04531.x

East Asian-type Helicobacter pylori cytotoxin-associated gene A protein has a more significant effect on growth of rat gastric mucosal cells than the Western type. / Fu, Hai Ying; Asahi, Kayoko; Hayashi, Yujiro et al.
In: Journal of Gastroenterology and Hepatology (Australia), Vol. 22, No. 3, 03.2007, p. 355-362.

Research output: Contribution to journal › Article › peer-review

Fu, HY, Asahi, K, Hayashi, Y, Eguchi, H, Murata, H, Tsujii, M, Tsuji, S, Azuma, T & Kawano, S 2007, 'East Asian-type Helicobacter pylori cytotoxin-associated gene A protein has a more significant effect on growth of rat gastric mucosal cells than the Western type', Journal of Gastroenterology and Hepatology (Australia), vol. 22, no. 3, pp. 355-362. https://doi.org/10.1111/j.1440-1746.2006.04531.x

@article{95ab4433378049e38c5c19c34b47c1ce,

title = "East Asian-type Helicobacter pylori cytotoxin-associated gene A protein has a more significant effect on growth of rat gastric mucosal cells than the Western type",

abstract = "Background and Aim: Cytotoxin-associated gene A (CagA) protein from H. pylori was reported to be injected into host gastric epithelial cells via a bacterial type IV secretion system, thereby modifying signal transduction. It is classified into two major subtypes, Western and East Asian. The present study aimed to compare the effects of East Asian-type and Western-type CagA on host cell growth. Methods: A tetracycline (tet)-off system and cagA genes from Western and East Asian-type H. pylori (NCTC 11637 and F32) were transfected into untransformed rat gastric mucosal (RGM1) cells to establish RGM1-CagA cell lines in which CagA expression could be controlled by tetracycline. These cell lines were used to investigate the effect of CagA protein expression on cell growth with BrdU and water-soluble tetrazolium reagent (WST-8) assays. CagA expression, phosphorylation and extracellular signal-regulated kinase (ERK) activation were examined with immunoprecipitation and Western blotting analysis. Results: 5-Bromo-2′deoxyuridine (BrdU) and WST-8 assays demonstrated significant increases in DNA replication and RGM1 cell growth after CagA protein expression. ERK phosphorylation was enhanced when CagA protein was expressed in RGM1-CagA cells. Moreover, the East Asian-type CagA showed a significantly greater effect on ERK activation and host cell growth than the Western type. PD98059, a mitogen-activated protein kinase kinase (MEK) inhibitor, suppressed ERK phosphorylation and the CagA protein-induced increase in RGM1-CagA cell growth. Conclusions: CagA protein expression induces an increase in RGM1-CagA cell proliferation via the mitogen-activated protein kinase (MAPK) signal pathway. The East Asian-type CagA showed a significantly greater effect on cell growth than the Western type, suggesting that the East Asian CagA-positive strain may have an important role in pathogenesis.",

keywords = "CagA, Cell growth, Helicobacter pylori, MAPK, RGM1 cells",

author = "Fu, {Hai Ying} and Kayoko Asahi and Yujiro Hayashi and Hiroshi Eguchi and Hiroaki Murata and Masahiko Tsujii and Shingo Tsuji and Takeshi Azuma and Sunao Kawano",

year = "2007",

month = mar,

doi = "10.1111/j.1440-1746.2006.04531.x",

language = "English (US)",

volume = "22",

pages = "355--362",

journal = "Journal of Gastroenterology and Hepatology (Australia)",

issn = "0815-9319",

publisher = "Wiley-Blackwell",

number = "3",

}

TY - JOUR

T1 - East Asian-type Helicobacter pylori cytotoxin-associated gene A protein has a more significant effect on growth of rat gastric mucosal cells than the Western type

AU - Fu, Hai Ying

AU - Asahi, Kayoko

AU - Hayashi, Yujiro

AU - Eguchi, Hiroshi

AU - Murata, Hiroaki

AU - Tsujii, Masahiko

AU - Tsuji, Shingo

AU - Azuma, Takeshi

AU - Kawano, Sunao

PY - 2007/3

Y1 - 2007/3

N2 - Background and Aim: Cytotoxin-associated gene A (CagA) protein from H. pylori was reported to be injected into host gastric epithelial cells via a bacterial type IV secretion system, thereby modifying signal transduction. It is classified into two major subtypes, Western and East Asian. The present study aimed to compare the effects of East Asian-type and Western-type CagA on host cell growth. Methods: A tetracycline (tet)-off system and cagA genes from Western and East Asian-type H. pylori (NCTC 11637 and F32) were transfected into untransformed rat gastric mucosal (RGM1) cells to establish RGM1-CagA cell lines in which CagA expression could be controlled by tetracycline. These cell lines were used to investigate the effect of CagA protein expression on cell growth with BrdU and water-soluble tetrazolium reagent (WST-8) assays. CagA expression, phosphorylation and extracellular signal-regulated kinase (ERK) activation were examined with immunoprecipitation and Western blotting analysis. Results: 5-Bromo-2′deoxyuridine (BrdU) and WST-8 assays demonstrated significant increases in DNA replication and RGM1 cell growth after CagA protein expression. ERK phosphorylation was enhanced when CagA protein was expressed in RGM1-CagA cells. Moreover, the East Asian-type CagA showed a significantly greater effect on ERK activation and host cell growth than the Western type. PD98059, a mitogen-activated protein kinase kinase (MEK) inhibitor, suppressed ERK phosphorylation and the CagA protein-induced increase in RGM1-CagA cell growth. Conclusions: CagA protein expression induces an increase in RGM1-CagA cell proliferation via the mitogen-activated protein kinase (MAPK) signal pathway. The East Asian-type CagA showed a significantly greater effect on cell growth than the Western type, suggesting that the East Asian CagA-positive strain may have an important role in pathogenesis.

AB - Background and Aim: Cytotoxin-associated gene A (CagA) protein from H. pylori was reported to be injected into host gastric epithelial cells via a bacterial type IV secretion system, thereby modifying signal transduction. It is classified into two major subtypes, Western and East Asian. The present study aimed to compare the effects of East Asian-type and Western-type CagA on host cell growth. Methods: A tetracycline (tet)-off system and cagA genes from Western and East Asian-type H. pylori (NCTC 11637 and F32) were transfected into untransformed rat gastric mucosal (RGM1) cells to establish RGM1-CagA cell lines in which CagA expression could be controlled by tetracycline. These cell lines were used to investigate the effect of CagA protein expression on cell growth with BrdU and water-soluble tetrazolium reagent (WST-8) assays. CagA expression, phosphorylation and extracellular signal-regulated kinase (ERK) activation were examined with immunoprecipitation and Western blotting analysis. Results: 5-Bromo-2′deoxyuridine (BrdU) and WST-8 assays demonstrated significant increases in DNA replication and RGM1 cell growth after CagA protein expression. ERK phosphorylation was enhanced when CagA protein was expressed in RGM1-CagA cells. Moreover, the East Asian-type CagA showed a significantly greater effect on ERK activation and host cell growth than the Western type. PD98059, a mitogen-activated protein kinase kinase (MEK) inhibitor, suppressed ERK phosphorylation and the CagA protein-induced increase in RGM1-CagA cell growth. Conclusions: CagA protein expression induces an increase in RGM1-CagA cell proliferation via the mitogen-activated protein kinase (MAPK) signal pathway. The East Asian-type CagA showed a significantly greater effect on cell growth than the Western type, suggesting that the East Asian CagA-positive strain may have an important role in pathogenesis.

KW - CagA

KW - Cell growth

KW - Helicobacter pylori

KW - MAPK

KW - RGM1 cells

UR - http://www.scopus.com/inward/record.url?scp=33846940534&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846940534&partnerID=8YFLogxK

U2 - 10.1111/j.1440-1746.2006.04531.x

DO - 10.1111/j.1440-1746.2006.04531.x

M3 - Article

AN - SCOPUS:33846940534

SN - 0815-9319

VL - 22

SP - 355

EP - 362

JO - Journal of Gastroenterology and Hepatology (Australia)

JF - Journal of Gastroenterology and Hepatology (Australia)

IS - 3

ER -

East Asian-type Helicobacter pylori cytotoxin-associated gene A protein has a more significant effect on growth of rat gastric mucosal cells than the Western type

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Cite this