Helicobacter pylori-secreting protein Tipα is a potent inducer of chemokine gene expressions in stomach cancer cells

  • Takashi Kuzuhara
  • Masami Suganuma
  • Miki Kurusu
  • Hirota Fujiki
Original Paper



Stomach cancer has a high mortality rate in East Asia, and is strongly associated with Helicobacter pylori (H. pylori) infection. H.pylori is known to express chemokine genes in the gastric mucosa, chemokines that are important host immune factors facilitating inflammation and tumor growth. To investigate the mechanism of carcinogenesis in the stomach, it is essential to determine which molecule of H. pylori is involved in induction of chemokines, but this has remained unclear. We previously reported that a tumor necrosis factor-α (TNF-α) inducing protein (Tipα) secreted from H. pylori acts as a tumor promoter in stomach cancer development, and thus started to investigate whether Tipα is involved in induction of chemokine genes.


Comprehensive gene expression analysis was conducted using DNA microarray and KeyMolnet analyses. The gene expression was quantitatively analyzed by real-time RT-PCR.


Comprehensive and quantitative gene expression analyses revealed that Tipα induces gene expression of the chemokines Ccl2, Ccl7, Ccl20, Cxcl1, Cxcl2, Cxcl5 and Cxcl10 extensively and simultaneously in mouse stomach cancer cells, MGT-40. Tipα induced high levels of chemokine gene expression, whereas inactive deleted Tipα, del-Tipα, showed only marginal expression, suggesting a correlation between tumor promotion and chemokine gene expression by Tipα. MG-132, a proteasome inhibitor which represses NF-κB-activation, inhibited chemokine gene expressions.


We report here that Tipα of H. pylori gene product is a strong inducer of chemokine gene expressions, providing a new model for stomach cancer development.


Tipα Helicobacter pylori Chemokine Stomach cancer Gastric epithelial cell 


  1. Balkwill F (2004) Cancer and the chemokine network. Nat Rev Cancer 4:540–550PubMedCrossRefGoogle Scholar
  2. De Valck D, Jin DY, Heyninck K, Van de Craen M, Contreras R, Fiers W, Jeang KT, Beyaert R (1999) The zinc finger protein A20 interacts with a novel anti-apoptotic protein which is cleaved by specific caspases. Oncogene 18:4182–4190PubMedCrossRefGoogle Scholar
  3. Eaton KA, Kersulyte D, Mefford M, Danon SJ, Krakowka S, Berg DE (2001) Role of Helicobacter pylori cag region genes in colonization and gastritis in two animal models. Infect Immun 69:2902–2908PubMedCrossRefGoogle Scholar
  4. Franco AT, Israel DA, Washington MK, Krishna U, Fox JG, Rogers AB, Neish AS, Collier-Hyams L, Perez-Perez GI, Hatakeyama M, Whitehead R, Gaus K, O’Brien DP, Romero-Gallo J, Peek RM Jr (2005) Activation of β-catenin by carcinogenic Helicobacter pylori. Proc Natl Acad Sci USA 102:10646–10651PubMedCrossRefGoogle Scholar
  5. Fujiki H, Suganuma M, Okabe S, Kurusu M, Imai K, Nakachi K (2002) Involvement of TNF-α changes in human cancer development, prevention and palliative care. Mech Ageing Dev 123:1655–1663PubMedCrossRefGoogle Scholar
  6. Haslett JN, Sanoudou D, Kho AT, Bennett RR, Greenberg SA, Kohane IS, Beggs AH, Kunkel LM (2002) Gene expression comparison of biopsies from duchenne muscular dystrophy (DMD) and normal skeletal muscle. Proc Natl Acad Sci USA 99:15000–15005PubMedCrossRefGoogle Scholar
  7. Hatakeyama M (2004) Oncogenic mechanisms of the Helicobacter pylori CagA protein. Nat Rev Cancer 4:688–694PubMedCrossRefGoogle Scholar
  8. Hu MC, Hung MC (2005) Role of IkappaB kinase in tumorigenesis. Future Oncol 1:67–78PubMedCrossRefGoogle Scholar
  9. IARC (1994) Working group on the evaluation of carcinogenic risks to humans. Lyon IARC 61:177–240Google Scholar
  10. Ichinose M, Nakanishi H, Fujino S, Tatematsu M (1998) Establishment and characterization of two cell lines from N-methyl-N-nitrosourea-induced mouse glandular stomach carcinomas. Jpn J Cancer Res 89:516–524PubMedGoogle Scholar
  11. Jin DY, Spencer F, Jeang KT (1998) Human T cell leukemia virus type 1 oncoprotein tax targets the human mitotic checkpoint protein MAD1. Cell 93:81–91PubMedCrossRefGoogle Scholar
  12. Kluver E, Adermann K, Schulz A (2006) Synthesis and structure-activity relationship of β-defensins, multi-functional peptides of the immune system. J Pept Sci 12:243–257PubMedCrossRefGoogle Scholar
  13. Liou HC, Hsia CY (2003) Distinctions between c-Rel and other NF-κB proteins in immunity and disease. Bioessays 25:767–780PubMedCrossRefGoogle Scholar
  14. Lu H, Wu JY, Kudo T, Ohno T, Graham DY, Yamaoka Y (2005) Regulation of interleukin-6 promoter activation in gastric epithelial cells infected with Helicobacter pylori. Mol Biol Cell 16:4954–4966PubMedCrossRefGoogle Scholar
  15. Nagasawa M, Kanzaki M, Iino Y, Morishita Y, Kojima I (2001) Identification of a novel chloride channel expressed in the endoplasmic reticulum, golgi apparatus, and nucleus. J Biol Chem 276:20413–20418PubMedCrossRefGoogle Scholar
  16. Peek Jr RM, Blaser MJ (2002) Helicobacter pylori and gastrointestinal tract adenocarcinoma. Nat Rev Cancer 2:28–37PubMedCrossRefGoogle Scholar
  17. Resnick MB, Sabo E, Meitner P, Kim SS, Cho Y, Kim H, Tavares R, Moss SF (2006) Global analysis of the human gastric epithelial transcriptome altered by H. pylori eradication in vivo. Gut May 24Google Scholar
  18. Richmond A (2002) NF-κB, chemokine gene transcription and tumour growth. Nat Rev Immunol 2:664–674PubMedCrossRefGoogle Scholar
  19. Sato H, Ishida S, Toda K, Matsuda R, Hayashi Y, Shigetaka M, Fukuda M, Wakamatsu Y, Itai A (2005) New approaches to mechanism analysis for drug discovery using DNA microarray data combined with KeyMolnet. Curr Drug Discov Technol 2:89–98PubMedCrossRefGoogle Scholar
  20. Sendera TJ, Dorris D, Ramakrishnan R, Nguyen A, Trakas D, Mazumder A (2002) Expression profiling with oligonucleotide arrays: technologies and applications for neurobiology. Neurochem Res 27:1005–1026PubMedCrossRefGoogle Scholar
  21. Suganuma M, Kurusu M, Okabe S, Sueoka N, Yoshida M, Wakatsuki Y, Fujiki H (2001) Helicobacter pylori membrane protein 1: a new carcinogenic factor of Helicobacter pylori. Cancer Res 61:6356–6359PubMedGoogle Scholar
  22. Suganuma M, Kurusu M, Suzuki K, Nishizono A, Murakami K, Fujioka T, Fujiki H (2005) New tumor necrosis factor-alpha-inducing protein released from Helicobacter pylori for gastric cancer progression. J Cancer Res Clin Oncol 131:305–313PubMedCrossRefGoogle Scholar
  23. Suganuma M, Kuzuhara T, Yamaguchi K, Fujiki H (2006) Carcinogenic role of tumor necrosis factor-alpha inducing protein of Helicobacter pylori in human stomach. J Biochem Mol Biol 39:1–8PubMedGoogle Scholar
  24. Toyoda H, Komurasaki T, Uchida D, Takayama Y, Isobe T, Okuyama T, Hanada K (1995) Epiregulin. A novel epidermal growth factor with mitogenic activity for rat primary hepatocytes. J Biol Chem 270:7495–7500PubMedCrossRefGoogle Scholar
  25. Visapaa I, Fellman V, Lanyi L, Peltonen L (2002) ABCB6 (MTABC3) excluded as the causative gene for the growth retardation syndrome with aminoaciduria, cholestasis, iron overload, and lactacidosis. Am J Med Genet 109:202–205PubMedCrossRefGoogle Scholar
  26. Wasmuth HE, Glantz A, Keppeler H, Simon E, Bartz C, Rath W, Mattsson LA, Marschall HU, Lammert F (2006) Intrahepatic cholestasis of pregnancy: the severe form is associated with common variants of the hepatobiliary phospholipid transporter gene ABCB4. Gut Aug 4Google Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Takashi Kuzuhara
    • 1
  • Masami Suganuma
    • 2
  • Miki Kurusu
    • 2
  • Hirota Fujiki
    • 1
  1. 1.Laboratory of Biochemistry, Faculty of Pharmaceutical SciencesTokushima Bunri UniversityTokushimaJapan
  2. 2.Saitama Cancer CenterResearch Institute for Clinical OncologySaitamaJapan

Personalised recommendations