Influence of Helicobacter pylori-derived outer membrane vesicles (OMVs) on Snail/β-Catenin cascade and metastasis-related proteins in 4T1 breast cancer cells
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Abstract
Background and Objectives: This study investigates the impact of Helicobacter pylori (H. pylori)-derived outer membrane vesicles (OMVs) on the regulation of Snail/β-Catenin cascade and the production of metastasis-related proteins, such as E-cadherin and Vimentin, in the 4T1 cell line.
Materials and Methods: OMVs were purified from H. pylori (ATCC 700392) cultures and applied to 4T1 cells at concentrations of 1, 5, and 10 μg/mL, with untreated cells serving as controls. The MTT assay was employed to quantify cell viability. Expression profiles of +vimentin, Snail, α-SMA, and β-catenin genes were evaluated by qRT-PCR, while protein expression of E-cadherin and Vimentin was analyzed via immunohistochemistry. Data were analyzed using appropriate statistical methods with SPSS and GraphPad Prism software.
Results: The MTT assay showed that 1 μg/mL OMVs were safe for normal cells. At this concentration, the expression of vimentin, Snail, α-SMA, and β-catenin genes significantly increased in the treatment group (P≤0.05). Additionally, Vimentin protein decreased, and E-cadherin protein increased (P≤0.05).
Conclusion: H. pylori-derived OMVs activate the Snail/β-Catenin cascade, influencing inflammatory responses and metastasis-related proteins, ultimately reducing migration in advanced cancer stages by modulating Vimentin and E-cadherin expression.
1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2021; 71: 209-249.
2. Gao X, Yang H, Chu Y, Zhang W, Wang Z, Ji L. The specific viral composition in triple-negative breast cancer tissue shapes the specific tumor microenvironment characterized on pathological images. Microb Pathog 2023; 184: 106385.
3. Smolarz B, Nowak AZ, Romanowicz H. Breast cancer-epidemiology, classification, pathogenesis and treatment (review of literature). Cancers (Basel) 2022; 14: 2569.
4. Yin L, Duan J-J, Bian X-W, Yu S-C. Triple-negative breast cancer molecular subtyping and treatment progress. Breast Cancer Res 2020; 22: 61.
5. Papaioannou NE, Beniata OV, Vitsos P, Tsitsilonis O, Samara P. Harnessing the immune system to improve cancer therapy. Ann Transl Med 2016; 4: 261.
6. Wang Y, Shi J, Chai K, Ying X, Zhou BP. The role of snail in EMT and tumorigenesis. Curr Cancer Drug Targets 2013; 13: 963-972.
7. Usman S, Waseem NH, Nguyen TKN, Mohsin S, Jamal A, Teh M-T, et al. Vimentin is at the heart of epithelial mesenchymal transition (EMT) mediated metastasis. Cancers (Basel) 2021; 13: 4985.
8. Song P, Gao Z, Bao Y, Chen L, Huang Y, Liu Y, et al. Wnt/β-catenin signaling pathway in carcinogenesis and cancer therapy. J Hematol Oncol 2024;17:46.
9. Akanda MR, Ahn E-J, Kim YJ, Salam SMA, Noh M-G, Lee T-K, et al. Analysis of stromal PDGFR-β and α-SMA expression and their clinical relevance in brain metastases of breast cancer patients. BMC Cancer 2023; 23: 468.
10. Rezaei Adriani R, Mousavi Gargari SL, Bakherad H, Amani J. Anti-EGFR bioengineered bacterial outer membrane vesicles as targeted immunotherapy candidate in triple-negative breast tumor murine model. Sci Rep 2023; 13: 16403.
11. Li Y, Wu J, Qiu X, Dong S, He J, Liu J, et al. Bacterial outer membrane vesicles-based therapeutic platform eradicates triple-negative breast tumor by combinational photodynamic/chemo-/immunotherapy. Bioact Mater 2022; 20: 548-560.
12. Furuyama N, Sircili MP. Outer membrane vesicles (OMVs) produced by gram-negative bacteria: structure, functions, biogenesis, and vaccine application. Biomed Res Int 2021; 2021: 1490732.
13. Zhang Z, Zhou Y, Zhao S, Ding L, Chen B, Chen Y. Nanomedicine‐enabled/augmented cell pyroptosis for efficient tumor nanotherapy. Adv Sci (Weinh) 2022; 9(35): e2203583.
14. González MF, Díaz P, Sandoval-Bórquez A, Herrera D, Quest AFG. Helicobacter pylori outer membrane vesicles and extracellular vesicles from Helicobacter pylori-infected cells in gastric disease development. Int J Mol Sci 2021; 22: 4823.
15. Murata-Kamiya N, Kurashima Y, Teishikata Y, Yamahashi Y, Saito Y, Higashi H, et al. Helicobacter pylori CagA interacts with E-cadherin and deregulates the β-catenin signal that promotes intestinal transdifferentiation in gastric epithelial cells. Oncogene 2007; 26: 4617-4626.
16. Baj J, Korona-Głowniak I, Forma A, Maani A, Sitarz E, Rahnama-Hezavah M, et al. Mechanisms of the epithelial–mesenchymal transition and tumor microenvironment in Helicobacter pylori-induced gastric cancer. Cells 2020; 9: 1055.
17. Shadi Vaziri S, Tajbakhsh E, Khamesipour F, Momtaz H, Mazaheri Z. Impact of Helicobacter pylori-Derived Outer Membrane Vesicles on Inflammation, Immune Responses, and Tumor Cell Migration in Breast Cancer Through the Snail/Β-Catenin Pathway. Rep Biochem Mol Biol 2024; 13: 263-272.
18. Brugge JS. Care and Passage of MCF-10A Cells In Monolayer Culture [Internet]. 2022. (https://brugge.hms.harvard.edu/system/files/protocols/Care_Passage_MCF_Matrigel.doc.pdf)
19. Qu Y, Han B, Yu Y, Yao W, Bose S, Karlan BY, et al. Evaluation of MCF10A as a reliable model for normal human mammary epithelial cells. PLoS One 2015; 10(7): e0131285.
20. Louis KS, Siegel AC. Cell viability analysis using trypan blue: manual and automated methods. Methods Mol Biol 2011; 740: 7-12.
21. Zoaiter M, Nasser R, Hage-Sleiman R, Abdel-Sater F, Badran B, Zeaiter Z. Helicobacter pylori outer membrane vesicles induce expression and secretion of oncostatin M in AGS gastric cancer cells. Braz J Microbiol 2021;52:1057-1066.
22. Yu H, Shen Y, Hong J, Xia Q, Zhou F, Liu X. The contribution of TGF-β in Epithelial-Mesenchymal Transition (EMT): Down-regulation of E-cadherin via snail. Neoplasma 2015; 62: 1-15.
23. Wei S, Li X, Wang J, Wang Y, Zhang C, Dai S, et al. Outer membrane vesicles secreted by Helicobacter pylori transmitting gastric pathogenic virulence factors. ACS Omega 2021; 7: 240-258.
24. Parker H, Chitcholtan K, Hampton MB, Keenan JI. Uptake of Helicobacter pylori outer membrane vesicles by gastric epithelial cells. Infect Immun 2010; 78: 5054-5061.
25. Bolori S, Shegefti S, Baghaei K, Yadegar A, Moon K-M, Foster LJ, et al. The effects of Helicobacter pylori‐derived outer membrane vesicles on hepatic stellate cell activation and liver fibrosis in vitro. Biomed Res Int 2023; 2023: 4848643.
26. Battaglia RA, Delic S, Herrmann H, Snider NT. Vimentin on the move: new developments in cell migration. F1000Res 2018; 7: F1000 Faculty Rev-1796.
27. Bruner HC, Derksen PWB. Loss of E-cadherin-dependent cell–cell adhesion and the development and progression of cancer. Cold Spring Harb Perspect Biol 2018; 10: a029330.
28. Jarzab M, Posselt G, Meisner-Kober N, Wessler S. Helicobacter pylori-derived outer membrane vesicles (OMVs): role in bacterial pathogenesis? Microorganisms 2020; 8: 1328.
29. Wu J, Li H, Xie H, Wu X, Lan P. The malignant role of exosomes in the communication among colorectal cancer cell, macrophage and microbiome. Carcinogenesis 2019; 40: 601-610.
30. Wu Z, Ma Q, Guo Y, You F. The role of Fusobacterium nucleatum in colorectal cancer cell proliferation and migration. Cancers (Basel) 2022; 14: 5350.
31. Shi L, Shangguan J, Lu Y, Rong J, Yang Q, Yang Y, et al. ROS-mediated up-regulation of SAE1 by Helicobacter pylori promotes human gastric tumor genesis and progression. J Transl Med 2024; 22: 148.
32. Ngo HKC, Lee HG, Piao JY, Zhong X, Lee HN, Han HJ, et al. Helicobacter pylori induces Snail expression through ROS‐mediated activation of Erk and inactivation of GSK‐3β in human gastric cancer cells. Mol Carcinog 2016; 55: 2236-2246.
33. Mobarez AM, Soleimani N, Esmaeili S-A, Farhangi B. Nanoparticle-based immunotherapy of breast cancer using recombinant Helicobacter pylori proteins. Eur J Pharm Biopharm 2020; 155: 69-76.
34. Liu Q, Li X, Zhang Y, Song Z, Li R, Ruan H, et al. Orally-administered outer-membrane vesicles from Helicobacter pylori reduce H. pylori infection via Th2-biased immune responses in mice. Pathog Dis 2019; 77: ftz050.
2. Gao X, Yang H, Chu Y, Zhang W, Wang Z, Ji L. The specific viral composition in triple-negative breast cancer tissue shapes the specific tumor microenvironment characterized on pathological images. Microb Pathog 2023; 184: 106385.
3. Smolarz B, Nowak AZ, Romanowicz H. Breast cancer-epidemiology, classification, pathogenesis and treatment (review of literature). Cancers (Basel) 2022; 14: 2569.
4. Yin L, Duan J-J, Bian X-W, Yu S-C. Triple-negative breast cancer molecular subtyping and treatment progress. Breast Cancer Res 2020; 22: 61.
5. Papaioannou NE, Beniata OV, Vitsos P, Tsitsilonis O, Samara P. Harnessing the immune system to improve cancer therapy. Ann Transl Med 2016; 4: 261.
6. Wang Y, Shi J, Chai K, Ying X, Zhou BP. The role of snail in EMT and tumorigenesis. Curr Cancer Drug Targets 2013; 13: 963-972.
7. Usman S, Waseem NH, Nguyen TKN, Mohsin S, Jamal A, Teh M-T, et al. Vimentin is at the heart of epithelial mesenchymal transition (EMT) mediated metastasis. Cancers (Basel) 2021; 13: 4985.
8. Song P, Gao Z, Bao Y, Chen L, Huang Y, Liu Y, et al. Wnt/β-catenin signaling pathway in carcinogenesis and cancer therapy. J Hematol Oncol 2024;17:46.
9. Akanda MR, Ahn E-J, Kim YJ, Salam SMA, Noh M-G, Lee T-K, et al. Analysis of stromal PDGFR-β and α-SMA expression and their clinical relevance in brain metastases of breast cancer patients. BMC Cancer 2023; 23: 468.
10. Rezaei Adriani R, Mousavi Gargari SL, Bakherad H, Amani J. Anti-EGFR bioengineered bacterial outer membrane vesicles as targeted immunotherapy candidate in triple-negative breast tumor murine model. Sci Rep 2023; 13: 16403.
11. Li Y, Wu J, Qiu X, Dong S, He J, Liu J, et al. Bacterial outer membrane vesicles-based therapeutic platform eradicates triple-negative breast tumor by combinational photodynamic/chemo-/immunotherapy. Bioact Mater 2022; 20: 548-560.
12. Furuyama N, Sircili MP. Outer membrane vesicles (OMVs) produced by gram-negative bacteria: structure, functions, biogenesis, and vaccine application. Biomed Res Int 2021; 2021: 1490732.
13. Zhang Z, Zhou Y, Zhao S, Ding L, Chen B, Chen Y. Nanomedicine‐enabled/augmented cell pyroptosis for efficient tumor nanotherapy. Adv Sci (Weinh) 2022; 9(35): e2203583.
14. González MF, Díaz P, Sandoval-Bórquez A, Herrera D, Quest AFG. Helicobacter pylori outer membrane vesicles and extracellular vesicles from Helicobacter pylori-infected cells in gastric disease development. Int J Mol Sci 2021; 22: 4823.
15. Murata-Kamiya N, Kurashima Y, Teishikata Y, Yamahashi Y, Saito Y, Higashi H, et al. Helicobacter pylori CagA interacts with E-cadherin and deregulates the β-catenin signal that promotes intestinal transdifferentiation in gastric epithelial cells. Oncogene 2007; 26: 4617-4626.
16. Baj J, Korona-Głowniak I, Forma A, Maani A, Sitarz E, Rahnama-Hezavah M, et al. Mechanisms of the epithelial–mesenchymal transition and tumor microenvironment in Helicobacter pylori-induced gastric cancer. Cells 2020; 9: 1055.
17. Shadi Vaziri S, Tajbakhsh E, Khamesipour F, Momtaz H, Mazaheri Z. Impact of Helicobacter pylori-Derived Outer Membrane Vesicles on Inflammation, Immune Responses, and Tumor Cell Migration in Breast Cancer Through the Snail/Β-Catenin Pathway. Rep Biochem Mol Biol 2024; 13: 263-272.
18. Brugge JS. Care and Passage of MCF-10A Cells In Monolayer Culture [Internet]. 2022. (https://brugge.hms.harvard.edu/system/files/protocols/Care_Passage_MCF_Matrigel.doc.pdf)
19. Qu Y, Han B, Yu Y, Yao W, Bose S, Karlan BY, et al. Evaluation of MCF10A as a reliable model for normal human mammary epithelial cells. PLoS One 2015; 10(7): e0131285.
20. Louis KS, Siegel AC. Cell viability analysis using trypan blue: manual and automated methods. Methods Mol Biol 2011; 740: 7-12.
21. Zoaiter M, Nasser R, Hage-Sleiman R, Abdel-Sater F, Badran B, Zeaiter Z. Helicobacter pylori outer membrane vesicles induce expression and secretion of oncostatin M in AGS gastric cancer cells. Braz J Microbiol 2021;52:1057-1066.
22. Yu H, Shen Y, Hong J, Xia Q, Zhou F, Liu X. The contribution of TGF-β in Epithelial-Mesenchymal Transition (EMT): Down-regulation of E-cadherin via snail. Neoplasma 2015; 62: 1-15.
23. Wei S, Li X, Wang J, Wang Y, Zhang C, Dai S, et al. Outer membrane vesicles secreted by Helicobacter pylori transmitting gastric pathogenic virulence factors. ACS Omega 2021; 7: 240-258.
24. Parker H, Chitcholtan K, Hampton MB, Keenan JI. Uptake of Helicobacter pylori outer membrane vesicles by gastric epithelial cells. Infect Immun 2010; 78: 5054-5061.
25. Bolori S, Shegefti S, Baghaei K, Yadegar A, Moon K-M, Foster LJ, et al. The effects of Helicobacter pylori‐derived outer membrane vesicles on hepatic stellate cell activation and liver fibrosis in vitro. Biomed Res Int 2023; 2023: 4848643.
26. Battaglia RA, Delic S, Herrmann H, Snider NT. Vimentin on the move: new developments in cell migration. F1000Res 2018; 7: F1000 Faculty Rev-1796.
27. Bruner HC, Derksen PWB. Loss of E-cadherin-dependent cell–cell adhesion and the development and progression of cancer. Cold Spring Harb Perspect Biol 2018; 10: a029330.
28. Jarzab M, Posselt G, Meisner-Kober N, Wessler S. Helicobacter pylori-derived outer membrane vesicles (OMVs): role in bacterial pathogenesis? Microorganisms 2020; 8: 1328.
29. Wu J, Li H, Xie H, Wu X, Lan P. The malignant role of exosomes in the communication among colorectal cancer cell, macrophage and microbiome. Carcinogenesis 2019; 40: 601-610.
30. Wu Z, Ma Q, Guo Y, You F. The role of Fusobacterium nucleatum in colorectal cancer cell proliferation and migration. Cancers (Basel) 2022; 14: 5350.
31. Shi L, Shangguan J, Lu Y, Rong J, Yang Q, Yang Y, et al. ROS-mediated up-regulation of SAE1 by Helicobacter pylori promotes human gastric tumor genesis and progression. J Transl Med 2024; 22: 148.
32. Ngo HKC, Lee HG, Piao JY, Zhong X, Lee HN, Han HJ, et al. Helicobacter pylori induces Snail expression through ROS‐mediated activation of Erk and inactivation of GSK‐3β in human gastric cancer cells. Mol Carcinog 2016; 55: 2236-2246.
33. Mobarez AM, Soleimani N, Esmaeili S-A, Farhangi B. Nanoparticle-based immunotherapy of breast cancer using recombinant Helicobacter pylori proteins. Eur J Pharm Biopharm 2020; 155: 69-76.
34. Liu Q, Li X, Zhang Y, Song Z, Li R, Ruan H, et al. Orally-administered outer-membrane vesicles from Helicobacter pylori reduce H. pylori infection via Th2-biased immune responses in mice. Pathog Dis 2019; 77: ftz050.
| Files | ||
| Issue | Vol 17 No 3 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i3.18824 | |
| Keywords | ||
| Breast neoplasms Helicobacter pylori Outer membrane vesicles Metastasis β-catenin Transcription factors | ||
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How to Cite
1.
Vaziri SS, Tajbakhsh E, Khamesipour F, Momtaz H. Influence of Helicobacter pylori-derived outer membrane vesicles (OMVs) on Snail/β-Catenin cascade and metastasis-related proteins in 4T1 breast cancer cells. Iran J Microbiol. 2025;17(3):411-419.
