Extracellular Membrane Vesicles of Escherichia coli Induce Apoptosis of CT26 Colon Carcinoma Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells Lines, Culture and Treatment
2.2. Reagents and Drugs
2.3. Extraction of E. coli-OMVs
2.4. Transmission Electron Microscopy
2.5. Nanoparticle Tracking Analysis
2.6. Labeling of E. coli-OMVs
2.7. Cells Viability Assay
2.8. Cell Invasion and Migration Analysis
2.9. Annexin V-FITC/PI Flow Cytometry Analysis
2.10. Mitochondrial Membrane Potential Assay
2.11. Western Blot
2.12. Xenograft Experiment In Vivo
2.13. Immunohistochemistry
2.14. Data Analysis
3. Results
3.1. Characterization of E. coli-OMVs
3.2. E. coli-OMVs Inhibit CT26 Colon Cancer Growth In Vivo
3.3. E. coli-OMVs Enter CT26 Cells and Inhibit Cell Viability
3.4. E. coli-OMVs Inhibited the Migration and Invasion of CT26 Cells
3.5. E. coli-OMVs Promoted Apoptosis of CT26 Cells
4. Discussion
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
- Boyle, P.; Langman, M.J. ABC of colorectal cancer: Epidemiology. BMJ 2000, 321, 0012452. [Google Scholar] [CrossRef]
- Aran, V.; Victorino, A.P.; Thuler, L.C.; Ferreira, C.G. Colorectal cancer: Epidemiology, disease mechanisms and interventions to reduce onset and mortality. Clin. Color. Cancer 2016, 15, 195–203. [Google Scholar] [CrossRef] [PubMed]
- El-Shami, K.; Oeffinger, K.C.; Erb, N.L.; Willis, A.; Bretsch, J.K.; Pratt-Chapman, M.L.; Cannady, R.S.; Wong, S.L.; Rose, J.; Barbour, A.L. American Cancer Society colorectal cancer survivorship care guidelines. CA A Cancer J. Clin. 2015, 65, 427–455. [Google Scholar] [CrossRef] [PubMed]
- Middlebrook, J.L.; Dorland, R.B. Bacterial toxins: Cellular mechanisms of action. Microbiol. Rev. 1984, 48, 199–221. [Google Scholar] [CrossRef]
- Ganai, S.; Arenas, R.B.; Sauer, J.P.; Bentley, B.; Forbes, N.S. In tumors Salmonella migrate away from vasculature toward the transition zone and induce apoptosis. Cancer Gene Ther. 2011, 18, 457–466. [Google Scholar] [CrossRef] [PubMed]
- Lee, C.; Lin, S.; Liu, J.; Chang, W.; Hsieh, J.; Wang, W. Salmonella induce autophagy in melanoma by the downregulation of AKT/mTOR pathway. Gene Ther. 2014, 21, 309–316. [Google Scholar] [CrossRef] [PubMed]
- Kim, S.H.; Castro, F.; Paterson, Y.; Gravekamp, C. High efficacy of a Listeria-based vaccine against metastatic breast cancer reveals a dual mode of action. Cancer Res. 2009, 69, 5860–5866. [Google Scholar] [CrossRef] [PubMed]
- Umer, B.; Good, D.; Anné, J.; Duan, W.; Wei, M.Q. Clostridial spores for cancer therapy: Targeting solid tumour microenvironment. J. Toxicol. 2012, 2012, 862764. [Google Scholar] [CrossRef] [PubMed]
- Kikuchi, Y.; Obana, N.; Toyofuku, M.; Kodera, N.; Soma, T.; Ando, T.; Fukumori, Y.; Nomura, N.; Taoka, A. Diversity of physical properties of bacterial extracellular membrane vesicles revealed through atomic force microscopy phase imaging. Nanoscale 2020, 12, 7950–7959. [Google Scholar] [CrossRef]
- Chen, Q.; Bai, H.; Wu, W.; Huang, G.; Li, Y.; Wu, M.; Tang, G.; Ping, Y. Bioengineering bacterial vesicle-coated polymeric nanomedicine for enhanced cancer immunotherapy and metastasis prevention. Nano Lett. 2019, 20, 11–21. [Google Scholar] [CrossRef]
- Chen, Y.; Liu, L.; Fu, H.; Wei, C.; Jin, Q. Comparative proteomic analysis of outer membrane vesicles from Shigella flexneri under different culture conditions. Biochem. Biophys. Res. Commun. 2014, 453, 696–702. [Google Scholar] [CrossRef] [PubMed]
- Jiang, Y.; Wang, L.; Yang, B.; Ma, G.; Chen, Z.; Ma, J.; Chang, X.; Fang, L.; Wang, Z. Bifidobacterium-derived membrane vesicles inhibit triple-negative breast cancer growth by inducing tumor cell apoptosis. Mol. Biol. Rep. 2023, 50, 7547–7556. [Google Scholar] [CrossRef]
- Shrivastava, S.; Jeengar, M.K.; Reddy, V.S.; Reddy, G.B.; Naidu, V. Anticancer effect of celastrol on human triple negative breast cancer: Possible involvement of oxidative stress, mitochondrial dysfunction, apoptosis and PI3K/Akt pathways. Exp. Mol. Pathol. 2015, 98, 313–327. [Google Scholar] [CrossRef] [PubMed]
- Carneiro, B.A.; El-Deiry, W.S. Targeting apoptosis in cancer therapy. Nat. Rev. Clin. Oncol. 2020, 17, 395–417. [Google Scholar] [CrossRef]
- Edlich, F. BCL-2 proteins and apoptosis: Recent insights and unknowns. Biochem. Biophys. Res. Commun. 2018, 500, 26–34. [Google Scholar] [CrossRef] [PubMed]
- Yaghoubi, A.; Khazaei, M.; Avan, A.; Hasanian, S.M.; Soleimanpour, S. The bacterial instrument as a promising therapy for colon cancer. Int. J. Color. Dis. 2020, 35, 595–606. [Google Scholar] [CrossRef]
- Toyofuku, M.; Nomura, N.; Eberl, L. Types and origins of bacterial membrane vesicles. Nat. Rev. Microbiol. 2019, 17, 13–24. [Google Scholar] [CrossRef]
- Kulp, A.; Kuehn, M.J. Biological functions and biogenesis of secreted bacterial outer membrane vesicles. Annu. Rev. Microbiol. 2010, 64, 163–184. [Google Scholar] [CrossRef]
- Schwechheimer, C.; Kuehn, M.J. Outer-membrane vesicles from Gram-negative bacteria: Biogenesis and functions. Nat. Rev. Microbiol. 2015, 13, 605–619. [Google Scholar] [CrossRef]
- Qing, S.; Lyu, C.; Zhu, L.; Pan, C.; Wang, S.; Li, F.; Wang, J.; Yue, H.; Gao, X.; Jia, R. Biomineralized bacterial outer membrane vesicles potentiate safe and efficient tumor microenvironment reprogramming for anticancer therapy. Adv. Mater. 2020, 32, 2002085. [Google Scholar] [CrossRef]
- Li, Y.; Zhao, R.; Cheng, K.; Zhang, K.; Wang, Y.; Zhang, Y.; Li, Y.; Liu, G.; Xu, J.; Xu, J. Bacterial outer membrane vesicles presenting programmed death 1 for improved cancer immunotherapy via immune activation and checkpoint inhibition. ACS Nano 2020, 14, 16698–16711. [Google Scholar] [CrossRef] [PubMed]
- Gujrati, V.; Kim, S.; Kim, S.-H.; Min, J.J.; Choy, H.E.; Kim, S.C.; Jon, S. Bioengineered bacterial outer membrane vesicles as cell-specific drug-delivery vehicles for cancer therapy. ACS Nano 2014, 8, 1525–1537. [Google Scholar] [CrossRef] [PubMed]
- Huang, W.; Shu, C.; Hua, L.; Zhao, Y.; Xie, H.; Qi, J.; Gao, F.; Gao, R.; Chen, Y.; Zhang, Q. Modified bacterial outer membrane vesicles induce autoantibodies for tumor therapy. Acta Biomater. 2020, 108, 300–312. [Google Scholar] [CrossRef] [PubMed]
- Kuerban, K.; Gao, X.; Zhang, H.; Liu, J.; Dong, M.; Wu, L.; Ye, R.; Feng, M.; Ye, L. Doxorubicin-loaded bacterial outer-membrane vesicles exert enhanced anti-tumor efficacy in non-small-cell lung cancer. Acta Pharm. Sin. B 2020, 10, 1534–1548. [Google Scholar] [CrossRef] [PubMed]
- Mattern, J.; Volm, M. Imbalance of cell proliferation and apoptosis during progression of lung carcinomas. Anticancer Res. 2004, 24, 4243–4246. [Google Scholar] [PubMed]
- Smets, L.A. Programmed cell death (apoptosis) and response to anti-cancer drugs. Anti-Cancer Drugs 1994, 5, 3–9. [Google Scholar] [CrossRef] [PubMed]
- Zimmermann, K.C.; Green, D.R. How cells die: Apoptosis pathways. J. Allergy Clin. Immunol. 2001, 108, S99–S103. [Google Scholar] [CrossRef] [PubMed]
- Korsmeyer, S.J.; Shutter, J.R.; Veis, D.J.; Merry, D.E.; Oltvai, Z.N. Bcl-2/Bax: A rheostat that regulates an anti-oxidant pathway and cell death. Semin. Cancer Biol. 1993, 4, 327–332. [Google Scholar] [PubMed]
- Joza, N.; Susin, S.A.; Daugas, E.; Stanford, W.L.; Cho, S.K.; Li, C.Y.; Sasaki, T.; Elia, A.J.; Cheng, H.-Y.M.; Ravagnan, L. Essential role of the mitochondrial apoptosis-inducing factor in programmed cell death. Nature 2001, 410, 549–554. [Google Scholar] [CrossRef]
- Lopez, J.; Tait, S. Mitochondrial apoptosis: Killing cancer using the enemy within. Br. J. Cancer 2015, 112, 957–962. [Google Scholar] [CrossRef]
- Abramowicz, I.; Carpenter, G.; Alfieri, M.; Colnaghi, R.; Outwin, E.; Parent, P.; Thauvin-Robinet, C.; Iaconis, D.; Franco, B.; O’Driscoll, M. Oral-facial-digital syndrome type I cells exhibit impaired DNA repair: Unanticipated consequences of defective OFD1 outside of the cilia network. Hum. Mol. Genet. 2017, 26, 19–32. [Google Scholar] [CrossRef] [PubMed]
- Srinivas, U.S.; Tan, B.W.; Vellayappan, B.A.; Jeyasekharan, A.D. ROS and the DNA damage response in cancer. Redox Biol. 2019, 25, 101084. [Google Scholar] [CrossRef] [PubMed]
- Zhao, Y.; Ye, X.; Xiong, Z.; Ihsan, A.; Ares, I.; Martínez, M.; Lopez-Torres, B.; Martínez-Larrañaga, M.-R.; Anadón, A.; Wang, X. Cancer metabolism: The role of ROS in DNA damage and induction of apoptosis in Cancer Cells. Metabolites 2023, 13, 796. [Google Scholar] [CrossRef] [PubMed]
- Norbury, C.J.; Zhivotovsky, B. DNA damage-induced apoptosis. Oncogene 2004, 23, 2797–2808. [Google Scholar] [CrossRef] [PubMed]
- O’Brien, K.; Breyne, K.; Ughetto, S.; Laurent, L.C.; Breakefield, X.O. RNA delivery by extracellular vesicles in mammalian cells and its applications. Nat. Rev. Mol. Cell Biol. 2020, 21, 585–606. [Google Scholar] [CrossRef] [PubMed]
- Bielaszewska, M.; Rüter, C.; Bauwens, A.; Greune, L.; Jarosch, K.-A.; Steil, D.; Zhang, W.; He, X.; Lloubes, R.; Fruth, A. Host cell interactions of outer membrane vesicle-associated virulence factors of enterohemorrhagic Escherichia coli O157: Intracellular delivery, trafficking and mechanisms of cell injury. PLoS Pathog. 2017, 13, e1006159. [Google Scholar] [CrossRef] [PubMed]
- Bielaszewska, M.; Rüter, C.; Kunsmann, L.; Greune, L.; Bauwens, A.; Zhang, W.; Kuczius, T.; Kim, K.S.; Mellmann, A.; Schmidt, M.A. Enterohemorrhagic Escherichia coli hemolysin employs outer membrane vesicles to target mitochondria and cause endothelial and epithelial apoptosis. PLoS Pathog. 2013, 9, e1003797. [Google Scholar] [CrossRef] [PubMed]
- Roberts, J.; Prager, M.D.; Bachynsky, N. The antitumor activity of Escherichia coli L-asparaginase. Cancer Res. 1966, 26, 2213–2217. [Google Scholar]
- Huang, X.; Wong, M.K.; Zhao, Q.; Zhu, Z.; Wang, K.Z.; Huang, N.; Ye, C.; Gorelik, E.; Li, M. Soluble recombinant endostatin purified from Escherichia coli: Antiangiogenic activity and antitumor effect. Cancer Res. 2001, 61, 478–481. [Google Scholar]
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Jiang, Y.; Ma, J.; Long, Y.; Dan, Y.; Fang, L.; Wang, Z. Extracellular Membrane Vesicles of Escherichia coli Induce Apoptosis of CT26 Colon Carcinoma Cells. Microorganisms 2024, 12, 1446. https://doi.org/10.3390/microorganisms12071446
Jiang Y, Ma J, Long Y, Dan Y, Fang L, Wang Z. Extracellular Membrane Vesicles of Escherichia coli Induce Apoptosis of CT26 Colon Carcinoma Cells. Microorganisms. 2024; 12(7):1446. https://doi.org/10.3390/microorganisms12071446
Chicago/Turabian StyleJiang, Yao, Jing Ma, Yuqing Long, Yuxi Dan, Liaoqiong Fang, and Zhibiao Wang. 2024. "Extracellular Membrane Vesicles of Escherichia coli Induce Apoptosis of CT26 Colon Carcinoma Cells" Microorganisms 12, no. 7: 1446. https://doi.org/10.3390/microorganisms12071446