Abstract：The aim of this study was to investigate the effect of outer membrane vesicles of Salmonella enterica serovar Typhi(S. Typhi-OMVs) on the proliferation of colorectal cancer cells under different culture conditions. In this study, ultracentrifugation was used to extract the outer membrane vesicles secreted by S. Typhi under the culture conditions of LB, adding acid, hyperosmotic, oxygen stress-derived LB and LPM, respectively. The morphology, particle size and protein distribution under the electron microscope were used to compare S. Typhi-OMVs cultured in different conditions.The colorectal cancer HT-29, SW480 and CT-26 cells were cultured in vitro. CCK-8 detected the effect of S. Typhi-OMVs (acid, hypertonic, oxygen stress and LPM culture conditions) at different time points on the proliferation of colorectal cancer cells, and the clone formation experiment was further verified. The cell cycle was detected by flow cytometry. To evaluate the drug toxicity of S. Typhi-OMVs in mice through body weight and HE staining sections of liver and kidney tissues.The results showed that there was no significant difference in morphology and size of S. Typhi-OMVs under different culture conditions. Compared with other culture conditions, S. Typhi could secrete more outer membrane vesicles under hyperosmotic stress. Under LB and acid stress, hyperosmotic and oxygen stress and LPM culture conditions, the protein distribution of S. Typhi-OMVs was 37~72KDa, 25~72KDa and 8 ~ 55 kDa respectively. S. Typhi-OMVs(LPM culture condition) can significantly inhibit the proliferation of SW480 and CT-26 cells, and arrest the cell cycle of SW480 cells in G2/M phase, and has no obvious hepatorenal toxicity to mice. It is concluded that S. Typhi-OMVs(LPM culture condition) had direct inhibitory effect on the proliferation of colorectal cancer cells (SW480, CT-26), and it was expected to be a therapeutic drug for colorectal cancer.
Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries[J]. CA Cancer J Clin,2021,71(3).
[3]
张宇涛.结直肠癌的治疗进展[J].山西医药杂志,2020,49(05):535-536.
[4]
van Zoggel D M G I,Bosman S J,Kusters M,Nieuwenhuijzen G A P,Cnossen J S,Creemers G J,van Lijnschoten G,Rutten H J T. Preliminary results of a cohort study of induction chemotherapy-based treatment for locally recurrent rectal cancer.[J]. Br J Surg,2018,105(4).
[5]
Schwechheimer C, Kuehn MJ. Outer-membrane vesicles from Gram-negative bacteria: biogenesis and functions.[J]. Nat Rev Microbiol,2015,13(10).
[6]
Bishop DG,Work E. An extracellular glycolipid produced by Escherichia coli grown under lysine-limiting conditions.[J]. Biochem J,1965,96(2).
[7]
Chatterjee SN,Das J. Electron microscopic observations on the excretion of cell-wall material by Vibrio cholerae.[J]. J Gen Microbiol,1967,49(1).
[8]
Sartorio MG, Pardue EJ, Feldman MF, Haurat MF. Bacterial Outer Membrane Vesicles: From Discovery to Applications.[J].Annu Rev Microbiol,2021,75.
[9]
Liang S, Wang C, Shao Y, Wang Y, Xing D, Geng Z. Recent advances in bacteria-mediated cancer therapy[J].Front Bioeng Biotechnol,2022,10.
Chen Q, Huang G, Wu W, Wang J, Hu J, Mao J, Chu PK, Bai H, Tang G. A Hybrid Eukaryotic-Prokaryotic Nanoplatform with Photothermal Modality for Enhanced Antitumor Vaccination.[J]. Adv Mater,2020,32(16).
[12]
Wang S, Guo J, Bai Y, Sun C, Wu Y, Liu Z, Liu X, Wang Y, Wang Z, Zhang Y, Hao H. Bacterial outer membrane vesicles as a candidate tumor vaccine platform [J].Front Immunol,2022,13.
[13]
Dell'Annunziata F, Folliero V, Giugliano R, De Filippis A, Santarcangelo C, Izzo V, Daglia M, Galdiero M, Arciola CR, Franci G. Gene Transfer Potential of Outer Membrane Vesicles of Gram-Negative Bacteria[J]. Int J Mol Sci,2021,22(11).
[14]
Baumgarten T, Sperling S, Seifert J, von Bergen M, Steiniger F, Wick LY, Heipieper HJ. Membrane vesicle formation as a multiple-stress response mechanism enhances Pseudomonas putida DOT-T1E cell surface hydrophobicity and biofilm formation.[J]. Appl Environ Microbiol,2012,78(17).
[15]
Bauwens A, Kunsmann L, Marejková M, Zhang W, Karch H, Bielaszewska M, Mellmann A. Intrahost milieu modulates production of outer membrane vesicles, vesicle-associated Shiga toxin 2a and cytotoxicity in Escherichia coli O157:H7 and O104:H4.[J].Environ Microbiol Rep,2017,9(5).
[16]
MacFadden DR, Bogoch II, Andrews JR. Advances in diagnosis, treatment, and prevention of invasive Salmonella infections.[J].Curr Opin Infect Dis,2016,29(5).
[17]
Mambu J, Virlogeux-Payant I, Holbert S, Grépinet O, Velge P, Wiedemann A. An Updated View on the Rck Invasin of Salmonella: Still Much to Discover[J].Front Cell Infect Microbiol,2017,7.
[18]
Kim OY, Park HT, Dinh NTH, Choi SJ, Lee J, Kim JH, Lee SW, Gho YS. Bacterial outer membrane vesicles suppress tumor by interferon-γ-mediated antitumor response.[J].Nat Commun,2017,8(1).
[19]
Kailoo S, Shreya, Kumar Y. Cytolethal distending toxin: from genotoxin to a potential biomarker and anti-tumor target.[J].World J Microbiol Biotechnol,2021,37(9).
