Skip to main content

Advertisement

SpringerLink
Log in

Cortactin Mediates Apoptosis of Gastric Epithelial Cells Induced by VacA Protein of Helicobacter pylori

  • Original Article
  • Published:
Digestive Diseases and Sciences Aims and scope Submit manuscript

Abstract

Background

Vacuolating cytotoxin antigen (VacA) is one of the major virulence factors in Helicobacter pylori (H. pylori), which is responsible for cell vacuolar degeneration and apoptotic cell death. A candidate host factor which mediates this process is cortactin, a protein associated with the processes of colonization and adhesion of H. pylori in gastric epithelium.

Aim

To investigate the role of cortactin in VacA-induced apoptosis of gastric epithelial cells.

Methods

Cortactin expression and shRNA lentiviral constructs were developed and transduced into the human gastric cancer cell line, AGS. VacA protein was purified from H. pylori cultures, acid-activated, and co-incubated with the transduced cell populations. Apoptosis was detected by flow cytometry, and the levels of the pro- and anti-apoptotic proteins Bax and Bcl-2 were determined by Western blot.

Results

Acid-activated purified VacA induced apoptosis in the parental AGS cells. Increased expression of cortactin (AGS/cortactin) led to a greater percentage of cells undergoing apoptosis. In contrast, knockdown of cortactin with shRNA (AGS/cortactin-shRNA) decreased the percentage of apoptotic cells. The protein levels of pro- and anti-apoptotic proteins Bax and Bcl-2 were increased and decreased in AGS/cortactin cells relative to the parental AGS cells. In the AGS/cortactin-shRNA cells, Bax protein levels were decreased, while Bcl-2 protein was increased.

Conclusions

The results indicate that cortactin is involved in the regulation of apoptosis induced by VacA in gastric cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  1. Aviles-Jimenez F, Reyes-Leon A, Nieto-Patlan E, et al. In vivo expression of Helicobacter pylori virulence genes in patients with gastritis, ulcer, and gastric cancer. Infect Immun. 2012;80:594–601.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  2. Blanchard TG, Czinn SJ, Correa P, et al. Genome sequences of 65 Helicobacter pylori strains isolated from asymptomatic individuals and patients with gastric cancer, peptic ulcer disease, or gastritis. Pathog Dis. 2013;68:39–43.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  3. Winter JA, Letley DP, Cook KW, et al. A role for the vacuolating cytotoxin, VacA, in colonization and Helicobacter pylori-induced metaplasia in the stomach. J Infect Dis. 2014;210:954–963.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Boquet P, Ricci V, Galmiche A, Gauthier NC. Gastric cell apoptosis and H. pylori: has the main function of VacA finally been identified? Trends Microbiol. 2003;11:410–413.

    Article  PubMed  CAS  Google Scholar 

  5. Manente L, Perna A, Buommino E, et al. The Helicobacter pylori’s protein VacA has direct effects on the regulation of cell cycle and apoptosis in gastric epithelial cells. J Cell Physiol. 2008;214:582–587.

    Article  PubMed  CAS  Google Scholar 

  6. Matsumoto A, Isomoto H, Nakayama M, et al. Helicobacter pylori VacA reduces the cellular expression of STAT3 and pro-survival Bcl-2 family proteins, Bcl-2 and Bcl-XL, leading to apoptosis in gastric epithelial cells. Dig Dis Sci. 2011;56:999–1006.

    Article  PubMed  CAS  Google Scholar 

  7. Zhao YQ, Guo T, Qian JM. Effects of broth culture filtrate protein of VacA+ Helicobacter pylori on the proliferation and apoptosis of gastric epithelial cells. Chin Med J (Engl). 2013;126:2168–2173.

    CAS  Google Scholar 

  8. Cho SJ, Kang NS, Park SY, Kim BO, Rhee DK, Pyo S. Induction of apoptosis and expression of apoptosis related genes in human epithelial carcinoma cells by Helicobacter pylori VacA toxin. Toxicon. 2003;42:601–611.

    Article  PubMed  CAS  Google Scholar 

  9. Roman-Roman A, Giono-Cerezo S, Camorlinga-Ponce M, Martinez-Carrillo DN, Loaiza-Loeza S, Fernandez-Tilapa G. VacA genotypes of Helicobacter pylori in the oral cavity and stomach of patients with chronic gastritis and gastric ulcer. Enferm Infecc Microbiol Clin. 2013;31:130–135.

    Article  PubMed  Google Scholar 

  10. Yahiro K, Satoh M, Nakano M, et al. Low-density lipoprotein receptor-related protein-1 (LRP1) mediates autophagy and apoptosis caused by Helicobacter pylori VacA. J Biol Chem. 2012;287:31104–31115.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  11. Ilatovskaya DV, Pavlov TS, Levchenko V, Negulyaev YA, Staruschenko A. Cortical actin binding protein cortactin mediates ENaC activity via Arp2/3 complex. FASEB J. 2011;25:2688–2699.

    Article  PubMed  CAS  Google Scholar 

  12. Khaleghian A, Takahashi A, et al. Cortactin, an actin binding protein, regulates GLUT4 translocation via actin filament remodeling. Biochemistry (Mosc). 2011;76:1262–1269.

    Article  Google Scholar 

  13. Cosen-Binker LI, Kapus A. Cortactin: the gray eminence of the cytoskeleton. Physiology (Bethesda). 2006;21:352–361.

    Article  CAS  Google Scholar 

  14. Luo C, Pan H, Mines M, Watson K, Zhang J, Fan GH. CXCL12 induces tyrosine phosphorylation of cortactin, which plays a role in CXC chemokine receptor 4-mediated extracellular signal-regulated kinase activation and chemotaxis. J Biol Chem. 2006;281:30081–30093.

    Article  PubMed  CAS  Google Scholar 

  15. Yang L, Kowalski JR, Yacono P, et al. Endothelial cell cortactin coordinates intercellular adhesion molecule-1 clustering and actin cytoskeleton remodeling during polymorphonuclear leukocyte adhesion and transmigration. J Immunol. 2006;177:6440–6449.

    Article  PubMed  CAS  Google Scholar 

  16. Noh SJ, Baek HA, Park HS, et al. Expression of SIRT1 and cortactin is associated with progression of non-small cell lung cancer. Pathol Res Pract. 2013;209:365–370.

    Article  PubMed  CAS  Google Scholar 

  17. Wang X, Cao W, Mo M, Wang W, Wu H, Wang J. VEGF and cortactin expression are independent predictors of tumor recurrence following curative resection of gastric cancer. J Surg Oncol. 2010;102:325–330.

    Article  PubMed  CAS  Google Scholar 

  18. Wei J, Zhao ZX, Li Y, Zhou ZQ, You TG. Cortactin expression confers a more malignant phenotype to gastric cancer SGC-7901 cells. World J Gastroenterol. 2014;20:3287–3300.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  19. Li X, Zheng H, Hara T, et al. Aberrant expression of cortactin and fascin are effective markers for pathogenesis, invasion, metastasis and prognosis of gastric carcinomas. Int J Oncol. 2008;33:69–79.

    PubMed  Google Scholar 

  20. Slanina H, Hebling S, Hauck CR, Schubert-Unkmeir A. Cell invasion by Neisseria meningitidis requires a functional interplay between the focal adhesion kinase, Src and cortactin. PLoS One. 2012;7:e39613.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  21. Selbach M, Moese S, Hurwitz R, Hauck CR, Meyer TF, Backert S. The Helicobacter pylori CagA protein induces cortactin dephosphorylation and actin rearrangement by c-Src inactivation. EMBO J. 2003;22:515–528.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  22. Tegtmeyer N, Wittelsberger R, Hartig R, Wessler S, Martinez-Quiles N, Backert S. Serine phosphorylation of cortactin controls focal adhesion kinase activity and cell scattering induced by Helicobacter pylori. Cell Host Microbe. 2011;9:520–531.

    Article  PubMed  CAS  Google Scholar 

  23. Adefolaju GA, Theron KE, Hosie MJ. Effects of HIV protease, nucleoside/non-nucleoside reverse transcriptase inhibitors on Bax, Bcl-2 and apoptosis in two cervical cell lines. Biomed Pharmacother. 2014;68:241–251.

    Article  PubMed  CAS  Google Scholar 

  24. Gauthier NC, Monzo P, Gonzalez T, et al. Early endosomes associated with dynamic F-actin structures are required for late trafficking of H. pylori VacA toxin. J Cell Biol. 2007;177:343–354.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  25. Hoshyar R, Bathaie SZ, Sadeghizadeh M. Crocin triggers the apoptosis through increasing the Bax/Bcl-2 ratio and caspase activation in human gastric adenocarcinoma, AGS, cells. DNA Cell Biol. 2013;32:50–57.

    Article  PubMed  CAS  Google Scholar 

  26. Samarghandian S, Nezhad MA, Mohammadi G. Role of caspases, Bax and Bcl-2 in chrysin-induced apoptosis in the A549 human lung adenocarcinoma epithelial cells. Anticancer Agents Med Chem. 2014;14:901–909.

    Article  PubMed  CAS  Google Scholar 

  27. de Bernard M, Papini E, de Filippis V, et al. Low pH activates the vacuolating toxin of Helicobacter pylori, which becomes acid and pepsin resistant. J Biol Chem. 1995;270:23937–23940.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This research was supported by the National Natural Science Fund (Grants 81171526 and 81472006) and the Chongqing Natural Science Fund Project (Grant CSTC 2011 jjA10023).

Author information

Author notes

    Authors and Affiliations

    1. Department of Gastroenterology, Daping Hospital, Third Military Medical University, 10 Changjiang Branch Road, Chongqing, 400042, China

      Hui Chang, Dongfeng Chen & Chunhui Lan

    2. Department of Microbiology and Biochemical Pharmacy, College of Pharmacy, The Third Military Medical University, Chongqing, China

      Bosheng Ni

    3. Cadre’s Ward of Bethune International Peace Hospital, PLA, Shijiazhuang City, Hebei Province, China

      Qianfei Zuo & Chunhua Wang

    4. Department of Gastroenterology, Guizhou Aerospace Hospital, Zunyi City, Guizhou Province, China

      Ran Han

    Authors
    1. Hui Chang
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Dongfeng Chen
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Bosheng Ni
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Qianfei Zuo
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Chunhua Wang
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Ran Han
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Chunhui Lan
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Chunhui Lan.

    Ethics declarations

    Conflict of interest

    The authors declare that they have no conflict of interest.

    Additional information

    Hui Chang and Dongfeng Chen have contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Chang, H., Chen, D., Ni, B. et al. Cortactin Mediates Apoptosis of Gastric Epithelial Cells Induced by VacA Protein of Helicobacter pylori . Dig Dis Sci 61, 80–90 (2016). https://doi.org/10.1007/s10620-015-3836-0

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s10620-015-3836-0

    Keywords

    Search

    Navigation