Investigation of bacterial abundance of Lactobacillus, Bifidobacterium, Enterococcus faecalis and Fusobacterium nucleatum in paraffin tissue samples of intestinal adenomatous polyps.
Prevalence of Lactobacillus, Bifidobacterium, Enterococcus faecalis and Fusobacterium nucleatum in intestinal adenomatous polyp tissue
Subject Areas : Molecular detection of biochemical and genetic markers
Mohamadreza Esrafili 1 , Reza Shapouri 2 , حبیب ضیغمی 3 , Fakhri Haghi 4
1 - Department of Microbiology,Faculty of Basic Sciences, Zanjan Branch,Islamic Azad University, Zanjan,Iran.
2 - Assistant Professor, Department of Microbiology, Zanjan Branch, Islamic Azad University, Zanjan, Iran
3 - دانشگاه علوم پزشکی زنجان
4 - Department of Microbiology and Virology, Zanjan University of Medical Sciences, Zanjan, Iran
Keywords: microbiome, adenomatous polyp, Lactobacillus, Bifidobacterium, Enterococcus faecalis, Fusobacterium nucleatum,
Abstract :
Background & Aim: Microbiota is a collection of microorganisms that live in the oral cavity, respiratory system and intestine of multicellular organisms. Microbiota exerts numerous physiological and pathological effects on the host in which it lives. Increasing attention has been directed to the interaction of host and microbiota. Adenomatous polyps are one of the common symptoms of colon cancer, the second leading cause of cancer-related death worldwide. Our study tries to show the relationship between Lactobacillus, Bifidobacterium, Enterococcus faecalis and Fusobacterium nucleatum in the intestinal paraffin tissue samples of adenomatous polyp patients and healthy individuals. Materials & methods: In this study, in order to investigate the mentioned bacteria in a total of 100 samples of intestinal paraffin tissue from adenomatous polyp patients (50 people) and healthy controls (50 people) for the presence, copy number and relative quantity of the above bacterial species using Real-time polymerase chain reaction (PCR), compared to the reference gene, were investigated. Results: In the studied samples, the presence and number of copies of Enterococcus faecalis bacteria in adenomatous polyp samples was significantly higher than the other three groups. There was no significant difference in the abundance and presence of Fusobacterium nucleatum and Lactobacillus species between the two groups. Also, a decrease in the average number of gaps and the relative abundance of Bifidobacterium species in adenomatous polyps compared to the control group was obtained. Conclusion: Our study showed a higher number of Enterococcus faecalis bacteria and a decrease in the number of Bifidobacterium species in the samples of intestinal paraffin tissue of patients with adenomatous polyps compared to the control group. However, any association between gut microbiome dysbiosis and adenomatous polyps remains unknown.
1. Backhed F, Ley RE, Sonnenburg JL, Peterson DA, Gordon JI. Host-bacterial mutualism in the human intestine. science. 2005 Mar 25;307(5717):1915-20.
2. Cukrowska B, Sowińska A, Bierła JB, Czarnowska E, Rybak A, Grzybowska-Chlebowczyk U. Intestinal epithelium, intraepithelial lymphocytes and the gut microbiota-Key players in the pathogenesis of celiac disease. World journal of gastroenterology. 2017 Nov 11;23(42):7505..
3. Hooper LV, Midtvedt T, Gordon JI. How host-microbial interactions shape the nutrient environment of the mammalian intestine. Annu Rev Nutr. 2002;22:283-307.
4. Ley RE, Turnbaugh PJ, Klein S, Gordon JI. Human gut microbes associated with obesity. nature. 2006 Dec 21;444(7122):1022-3.
5. Schiffman M, Castle PE, Jeronimo J, Rodriguez AC, Wacholder S. Human papillomavirus and cervical cancer. Lancet. 2007; 370:890–907. doi: 10.1016/S0140- 6736(07)61416-0.
6. Zeller G, Tap J, Voigt AY, Sunagawa S, Kultima JR, Costea PI, Amiot A, Böhm J, Brunetti F, Habermann N, Hercog R. Potential of fecal microbiota for early‐stage detection of colorectal cancer. Molecular systems biology. 2014 Nov;10(11):766.
7. Øines M, Helsingen LM, Bretthauer M, Emilsson L. Epidemiology and risk factors of colorectal polyps. Best Practice & Research Clinical Gastroenterology. 2017 Aug 1;31(4):419-24.
8. -8 Pan J, Cen L, Xu L, Miao M, Li Y, Yu C, Shen Z. Prevalence and risk factors for colorectal polyps in a Chinese population: a retrospective study. Scientific Reports. 2020 Apr 24;10(1):6974.-
9. Thursby, E.; Juge, N. Introduction to the human gut microbiota. Biochem. J. 2017, 474, 1823–1836.
10. Qin J, Li R, Raes J, Arumugam M, Burgdorf KS, Manichanh C, Nielsen T, Pons N, Levenez F, Yamada T, Mende DR. A human gut microbial gene catalogue established by metagenomic sequencing. nature. 2010 Mar 4;464(7285):59-65..
11. Ferlay, J.; Steliarova-Foucher, E.; Lortet-Tieulent, J.; Rosso, S.; Coebergh, J.; Comber, H.; Forman, D.; Bray, F. Cancer incidence and mortality patterns in Europe: Estimates for 40 countries in 2012. Eur. J. Cancer 2013, 49, 1374–1403.
12. Larsson, S.C.; Wolk, A. Meat consumption and risk of colorectal cancer: A meta-analysis of prospective studies. Int. J. Cancer 2006, 119, 2657–2664.
13. Dahm, C.C.; Keogh, R.H.; Spencer, E.A.; Greenwood, D.C.; Key, T.J.; Fentiman, I.S.; Shipley, M.J.; Brunner, E.J.; Cade, J.E.; Burley, V.J.; et al. Dietary Fiber and Colorectal Cancer Risk: A Nested Case-Control Study Using Food Diaries. J. Natl. Cancer Inst. 2010, 102, 614–626.
14. Liang, S.Y.; Mao, Y.; Liao, M.; Xu, Y.S.; Chen, Y.C.; Huang, X.L.; Wei, C.Y.; Wu, C.T.; Wang, Q.Y.; Pan, X.Y.; et al. Gut microbiome associated with APC gene mutation in patients with intestinal adenomatous polyps. Int. J. Biol. Sci. 2020, 16, 135–146.
15. Nosho, K.; Sukawa, Y.; Adachi, Y.; Ito, M.; Mitsuhashi, K.; Kurihara, H.; Kanno, S.; Yamamoto, I.; Ishigami, K.; Igarashi, H.; et al. Association of Fusobacterium nucleatum with immunity and molecular alterations in colorectal cancer. World J. Gastroenterol. 2016, 22, 557.
16. Stanton C, Gardiner G, Meehan H, Collins K, Fitzgerald G, Lynch PB, et al. Market potential for probiotics. Am J Clin Nutr. 2001;73(Suppl 2):476S-83S.
17. De Roos NM, Katan MB. Effects of probiotic bacteria on diarrhea, lipid metabolism, and carcinogenesis: a review of papers published between 1988 and 1998. The American journal of clinical nutrition. 2000 Feb 1;71(2):405-11..
18. Russo F, Linsalata M, Orlando A. Probiotics against neoplastic transformation of gastric mucosa: effects on cell proliferation and polyamine metabolism. World J Gastroenterol. 2014;20:13258-72.
19. Bundgaard-Nielsen C, Baandrup UT, Nielsen LP, Sorensen S. The presence of bacteria varies between colorectal adenocarcinomas, precursor lesions and non-malignant tissue. BMC Cancer. 2019;19(1):399
20. Huijsmans CJ, Damen J, van der Linden JC, Savelkoul PH, Hermans MH. Comparative analysis of four methods to extract DNA from paraffin-embedded tissues: effect on downstream molecular applications. BMC research notes. 2010 Dec;3:1-9.
21. Jukes TH, Cantor CR. Evolution of protein molecules. Mammalian protein metabolism. 1969;3(24):21-132.
22. -Ritchie LE, Steiner JM, Suchodolski JS. Assessment of microbial diversity along the feline intestinal tract using 16S rRNA gene analysis. FEMS microbiology ecology. 2008 Dec 1;66(3):590-8.
23. Cannon K, Byrne B, Happe J, Wu K, Ward L, Chesnel L, et al. Enteric microbiome profiles during a randomized phase 2 clinical trial of surotomycin versus vancomycin for the treatment of Clostridium difficile infection. J Antimicrob Chemother. 2017;72(12):3453–61.
24. Grady WM, Markowitz SD. The molecular pathogenesis of colorectal cancer and its potential application to colorectal cancer screening. Digestive diseases and sciences. 2015 Mar;60:762-72.
25. Zeller G, Tap J, Voigt AY, Sunagawa S, Kultima JR, Costea PI, Amiot A, Böhm J, Brunetti F, Habermann N, Hercog R. Potential of fecal microbiota for early‐stage detection of colorectal cancer. Molecular systems biology. 2014 Nov;10(11):766.
26. Rezasoltani S, Asadzadeh-Aghdaei H, Nazemalhosseini-Mojarad E, Dabiri H, Ghanbari R, Zali MR. Gut microbiota, epigenetic modification and colorectal cancer. Iranian journal of microbiology. 2017 Apr;9(2):55.
27. Peters BA, Dominianni C, Shapiro JA, Church TR, Wu J, Miller G, Yuen E, Freiman H, Lustbader I, Salik J, Friedlander C. The gut microbiota in conventional and serrated precursors of colorectal cancer. Microbiome. 2016 Dec;4:1-4.
28. Balamurugan R, Rajendiran E, George S, Samuel GV, Ramakrishna BS. Real-time polymerase chain reaction quantification of specific butyrateproducing bacteria, Desulfovibrio and enterococcus faecalis in the feces of patients with colorectal cancer. J Gastroenterol Hepatol. 2008;23(8 Pt1):1298–303.
29. Niya MHK, Basi A, Koochak A, Tameshkel FS, Rakhshani N, Zamani F, et al. Sensitive high-resolution melting analysis for screening of KRAS and BRAF mutations in Iranian human metastatic colorectal cancers. Asian Pac J Cancer Prev. 2016;17:5147.
30. Zeighami H , Khodaverdi N , Jalilvand A, Haghi F . High frequency of enterotoxigenic Bacteroides fragilis and Enterococcus faecalis in the paraffin-embedded tissues of Iranian colorectal cancer patients. J BMC Cancer (2021) 21:1353.
31. 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.
32. Zhou Y, He H, Xu H, Li Y, Li Z, Du Y, He J, Zhou Y, Wang H, Nie Y. Association of oncogenic bacteria with colorectal cancer in South China. Oncotarget. 2016 Dec 12;7(49):80794.
33. Kashani N, Abadi AB, Rahimi F, Forootan M. FadA-positive Fusobacterium nucleatum is prevalent in biopsy specimens of Iranian patients with colorectal cancer.
New microbes and new infections. 2020 Mar 1;34:100651.
34. Rezasoltani S, Aghdaei HA, Dabiri H, Sepahi AA, Modarressi MH, Mojarad EN. The association between fecal microbiota and different types of colorectal polyp as precursors of colorectal cancer. Microbial pathogenesis. 2018 Nov 1;124:244-9.
35. Kashani N, Abadi AB, Rahimi F, Forootan M. FadA-positive Fusobacterium nucleatum is prevalent in biopsy specimens of Iranian patients with colorectal cancer. New microbes and new infections. 2020 Mar 1;34:100651.
36. Repass J. Replication Study: Fusobacterium nucleatum infection is prevalent in human colorectal carcinoma. Elife. 2018 Mar 13;7:e25801..
37. Gorkiewicz G, Moschen A. Gut microbiome: a new player in gastrointestinal disease. Virchows Archiv. 2018 Jan;472(1):159-72.
38. Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, Morelli L, Canani RB, Flint HJ, Salminen S, et al. Expert consensus document. The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol. 2014;11:506–14.
39. -39 Tiptiri-Kourpeti A, Spyridopoulou K, Santarmaki V, Aindelis G, Tompoulidou E, Lamprianidou EE, Saxami G, Ypsilantis P, Lampri ES, Simopoulos C, et al. Lactobacillus casei exerts anti-proliferative effects accompanied by apoptotic cell death and up-regulation of TRAIL in colon carcinoma cells. PLoS ONE. 2016;11:e0147960.
40. Abrahamse SL, Pool-Zobel BL, Rechkemmer G. Potential of short chain fatty acids to modulate the induction of DNA damage and changes in the intracellular calcium concentration by oxidative stress in isolated rat distal colon cells. Carcinogenesis. 1999 Apr 1;20(4):629-34.
41. Wang T, Cai G, Qiu Y, Fei N, Zhang M, Pang X, Jia W, Cai S, Zhao L. Structural segregation of gut microbiota between colorectal cancer patients and healthy volunteers. The ISME journal. 2012 Feb;6(2):320-9.
42. Davis CD, Milner JA. Gastrointestinal microflora, food components and colon cancer prevention. The Journal of nutritional biochemistry. 2009 Oct 1;20(10):743-52.
43. Rezasoltani S, Asadzadeh-Aghdaei H, Nazemalhosseini-Mojarad E, Dabiri H, Ghanbari R, Zali MR. Gut microbiota, epigenetic modification and colorectal cancer. Iranian journal of microbiology. 2017 Apr;9(2):55.