The effect of intestinal microbiota metabolites on HT29 cell line using MTT method in patients with colorectal cancer
Gastroenterology and Hepatology from Bed to Bench,
Aim: The aim of this study was to evaluate the effect of intestinal microbiota metabolites in colorectal cancer patients on HT29 cell line using MTT assay.
Background: Colorectal cancer is one of the most common malignant tumors. Human guts harbor abundant microbes that adjust many aspects of the host physiology. Increasing studies suggest that gut microbiota play a significant role in the incidence and expansion of CRC, as a result of virulence factors, bacterial metabolites, or inflammatory pathways.
Methods: In this cross-sectional study, 60 biopsy samples including 30 cancerous and 30 adjacent healthy tissues were collected from patients with CRC during 2017. Biopsy samples were first cultured on Thioglycollate broth medium for 24hr after which the microbiota metabolites were filtered and stored at -20 C° for further evaluation. HT29 cells were treated by microbiota metabolites at different times (3, 6, 12, 18h) and its viability was assessed by MTT assay.
Results: The cells treated with microbiota metabolites showed increased viability and proliferation in time-dependent analysis by MTT assay, but there was not significant differences between the two groups.
Conclusion: It seems that microbial metabolites are able to induce proliferation and increase cell viability and thus induce colorectal cancer.
Keywords: Metabolites, Cell line, MTT, Colorectal cancer.
(Please cite as: Jahani-Sherafat S, Azimirad M, Ghasemian-Safaei H, Ahmadi Amoli H, Moghim SH, Sherkat Gh, et al. The effect of intestinal microbiota metabolites on HT29 cell line using MTT method in patients with colorectal cancer. Gastroenterol Hepatol Bed Bench 2019;12(Suppl.1):S74-S79).
- Cell line
- Colorectal cancer.
How to Cite
Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 2015 Mar 1;136(5):E359-86.
Dolatkhah R, Somi MH, Bonyadi MJ, Asvadi Kermani I, Farassati F, Dastgiri S. Colorectal cancer in iran: molecular epidemiology and screening strategies. J Cancer Epidemiol. 2015;2015:643020.
Ansari R, Mahdavinia M, Sadjadi A, Nouraie M, Kamangar F, Bishehsari F, et al. Incidence and age distribution of colorectal cancer in Iran: results of a population-based cancer registry. Cancer Lett. 2006;240(1):143-7.
Serban DE. The gut microbiota in the metagenomics era: sometimes a friend, sometimes a foe. Roumanian archives of microbiology and immunology. 2011 Jul-Sep;70(3):134-40.
Uronis JM, Mühlbauer M, Herfarth HH, Rubinas TC, Jones GS, Jobin C. Modulation of the intestinal microbiota alters colitis-associated colorectal cancer susceptibility. PLoS One. 2009 Jun 24;4(6):e6026
Sharma R, Young C, Mshvildadze M, Neu J. Intestinal Microbiota Does It Play a Role in Diseases of the Neonate? Neoreviews. 2009;10(4):e166-e79.
Marchesi JR, Dutilh BE, Hall N, Peters W, Roelofs R, Boleij A, et al. Towards the human colorectal cancer microbiome. PloS one. 2011;6(5):e20447.
Prakash S, Rodes L, Coussa-Charley M, Tomaro-Duchesneau C. Gut microbiota: next frontier in understanding human health and development of biotherapeutics. Biologics. 2011;5:71-86.
Ridlon JM, Kang D-J, Hylemon PB. Bile salt biotransformations by human intestinal bacteria. J Lipid Res. 2006;47(2):241-59.
Dabek M, McCrae SI, Stevens VJ, Duncan SH, Louis P. Distribution of β-glucosidase and β-glucuronidase activity and of β-glucuronidase gene gus in human colonic bacteria. FEMS Microbiol Ecol. 2008;66(3):487-95.
Kim D-H, Jin Y-H. Intestinal bacterial β-glucuronidase activity of patients with colon cancer. Arch Pharm Res. 2001;24(6):564-7.
Wang R-F, Chen H, Paine DD, Cerniglia CE. Microarray method to monitor 40 intestinal bacterial species in the study of azo dye reduction. Biosens Bioelectron.2004;20(4):699-705.
Carroll IM, Maharshak N. Enteric bacterial proteases in inflammatory bowel disease-pathophysiology and clinical implications. World J of Gastroenterol.2013;19(43):7531.
Tahara T, Yamamoto E, Suzuki H, Maruyama R, Chung W, Garriga J, et al. Fusobacterium in colonic flora and molecular features of colorectal carcinoma. Cancer Res. 2014;74(5):1311-8
Viljoen KS, Dakshinamurthy A, Goldberg P, Blackburn JM. Quantitative Profiling of Colorectal Cancer-Associated Bacteria Reveals Associations between Fusobacterium spp., Enterotoxigenic Bacteroides fragilis (ETBF) and Clinicopathological Features of Colorectal Cancer. PloS one. 2015;10(3):e0119462.
Sadeghi Ekbatan Sh, Li XQ, Ghorbani M, Azadi B, Kubow S. Chlorogenic Acid and Its Microbial Metabolites Exert Anti-Proliferative Effects, S-Phase Cell-Cycle Arrest and Apoptosis in Human Colon Cancer Caco-2 Cells. Int J Mol Sci. 2018; 19(3): pii: E723
Green RA, Kaplan KB. Chromosome instability in colorectal tumor cells is associated with defects in microtubule plus-end attachments caused by a dominant mutation in APC. The Journal of cell biology. 2003;163(5):949-61.
Francescone R, Hou V, Grivennikov SI. Microbiome, inflammation and cancer. Cancer J. 2014 May-Jun; 20(3): 181–189.
Blumberg R, Powrie F. Microbiota, disease, and back to health: a metastable journey. Sci Transl Med. 2012 Jun 6;4(137):137rv7.
Tsilimigras MC, Fodor A, Jobin C. Carcinogenesis and therapeutics: the microbiota perspective. Nat Microbiol. 2017;2(3):17008.
Cani PD, Plovier H, Van Hul M, Geurts L, Delzenne NM, Druart C, et al. Endocannabinoids—at the crossroads between the gut microbiota and host metabolism. Nat Rev Endocrinol. 2016;12(3):133.
Turroni F, Peano C, Pass DA, Foroni E, Severgnini M, Claesson MJ, et al. Diversity of bifidobacteria within the infant gut microbiota. PLoS One. 2012;7(5):e36957.
Carman RJ, Simon MA, Fernández H, Miller MA, Bartholomew MJ. Ciprofloxacin at low levels disrupts colonization resistance of human fecal microflora growing in chemostats. Regul Toxicol Pharmacol. 2004 Dec;40(3):319-26.
Shi H, Guo Y, Liu Y, Binlin Shi, Guo X, Jin L, Yan S, The in vitro effect of lipopolysaccharide on proliferation, inflammatory factors and antioxidant enzyme activity in bovine mammary epithelial cells. Animal Nutrition, 2016; 99-104
Wang DX. A research review: poisonous bacteria lipopolysaccharide and development Foreign Med Sci Hyg. 2008; 35 (2008):170- 173
Venturi M, Hambly R J, Glinghammar B, Rafter J J, Rowland I R. Genotoxic activity in human faecal water and the role of bile acids: a study using the alkaline comet assay. Carcinogenesis. 1997 Dec;18(12):2353-9.
Tilg H, Adolph T E, Gerner R R, Moschen A R. The Intestinal Microbiota in Colorectal Cancer. Cancer Cell. 2018 Jun 11;33(6):954-964
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