Intestinal bacteria detected in cancer and adjacent tissue from patients with colorectal cancer

Chen-Jian Liu, Yuan-Lian Zhang, Yun Shang, Bian Wu, En Yang, Yi-Yong Luo, Xiao-Ran Li, Chen-Jian Liu, Yuan-Lian Zhang, Yun Shang, Bian Wu, En Yang, Yi-Yong Luo, Xiao-Ran Li

Abstract

Intestinal bacteria are symbiotic microbiota within the human gut and are implicated in the occurrence and development of colorectal cancer (CRC). The current study investigated the changes in bacterial composition prior to and following surgery, as well as the differences in the bacterial community structure between cancer tissue and adjacent normal tissue. The diversity of the bacterial community and the composition of the bacteria were assessed. In addition, phylogenetic analysis and principle component analysis (PCA) were performed. The results revealed that cancer tissue and adjacent normal tissue exhibited similar bacterial compositions. However, a significant difference was identified in the composition of intestinal bacteria in stool samples collected from patients following surgery compared with stool samples collected prior to surgery. Each patient had their own unique intestinal bacterial community, likely due to a number of factors, including diet, genetic factors and health status. In addition, phylogenetic trees revealed that the most abundant operational taxonomic unit, 0001, was associated with Escherichia coli in all samples. Finally, PCA suggested that the bacterial community structure in all patient stools was similar following surgery. The current study provides information regarding the diversity of the intestinal bacterial community of patients with CRC and provides a basis for postoperative intestinal assessments.

Keywords: bacterial composition; colorectal cancer; stool; surgery; tissue.

Figures

Figure 1.
Figure 1.
Bacterial composition of all stool samples obtained from patients prior to and following surgery. The bacterial community structures of stool samples were analyzed at the (A) phylum level, (B) family level and (C) genus level.
Figure 2.
Figure 2.
Bacterial compositions of stool samples obtained from each patient prior to and following surgery. (A-H) The bacterial composition of stool samples from every patient was analyzed at the genus level.
Figure 3.
Figure 3.
Bacterial composition of all tissue samples obtained from the patients. All sequences were analyzed at the (A) phylum level, (B) family level and (C) genus level. C, cancerous tissue; N, adjacent normal tissue.
Figure 4.
Figure 4.
Bacterial compositions of all tissue samples obtained from each patient. (A-H) All sequences of every patient were analyzed at the genus level. C, cancerous tissue; N, adjacent normal tissue.
Figure 5.
Figure 5.
Phylogenetic tree of 16S ribosomal RNA gene sequences based on a cut-off value of 0.03. A total of 44 OTUs were used to construct a phylogenetic tree. The OTU names were based on the number of sequences, therefore, the lower the number, the higher its abundance. OTU, operational taxonomic unit.
Figure 6.
Figure 6.
PCA based on different sample sources and different patients. PCA was performed based on (A) different sample sources, including cancer tissue, adjacent normal tissue and stool samples obtained prior to and following surgery, and (B) different patients. PCA, principle component analysis; PC, principle component.

References

    1. Bäckhed F, Ley RE, Sonnenburg JL, Peterson DA, Gordon JI. Host-bacterial mutualism in the human intestine. Science. 2005;307:1915–1920. doi: 10.1126/science.1104816.
    1. Qin J, Li R, Raes J, Arumugam M, Burgdorf KS, Manichanh C, Nielsen T, Pons N, Levenez F, Yamada T, et al. A human gut microbial gene catalogue established by metagenomic sequencing. Nature. 2010;464:59–65. doi: 10.1038/nature08821.
    1. Gao Z, Guo B, Gao R, Zhu Q, Qin H. Microbiota disbiosis is associated with colorectal cancer. Front Microbiol. 2015;6:20. doi: 10.3389/fmicb.2015.00020.
    1. Sobhani I, Tap J, Roudot-Thoraval F, Roperch JP, Letulle S, Langella P, Corthier G, Van Nhieu Tran J, Furet JP. Microbial dysbiosis in colorectal cancer (CRC) patients. PLoS One. 2011;6:e16393. doi: 10.1371/journal.pone.0016393.
    1. Larsen N, Vogensen FK, van den Berg FW, Nielsen DS, Andreasen AS, Pedersen BK, Al-Soud WA, Sørensen SJ, Hansen LH, Jakobsen M. Gut microbiota in human adults with type 2 diabetes differs from non-diabetic adults. PLoS One. 2010;5:e9085. doi: 10.1371/journal.pone.0009085.
    1. Tannock GW. Molecular analysis of the intestinal microflora in IBD. Mucosal Immunol. 2008;1(Suppl 1):S15–S18. doi: 10.1038/mi.2008.54.
    1. Ley RE, Turnbaugh PJ, Klein S, Gordon JI. Microbial ecology: Human gut microbes associated with obesity. Nature. 2006;444:1022–1023. doi: 10.1038/4441022a.
    1. Brenner H, Kloor M, Pox CP. Colorectal cancer. Lancet. 2014;383:1490–1502. doi: 10.1016/S0140-6736(13)61649-9.
    1. Burt RW. Colon cancer screening. Gastroenterology. 2000;119:837–853. doi: 10.1053/gast.2000.16508.
    1. Platz EA, Willett WC, Colditz GA, Rimm EB, Spiegelman D, Giovannucci E. Proportion of colon cancer risk that might be preventable in a cohort of middle-aged US men. Cancer Causes Control. 2000;11:579–588. doi: 10.1023/A:1008999232442.
    1. Watson AJ, Collins PD. Colon cancer: A civilization disorder. Dig Dis. 2011;29:222–228. doi: 10.1159/000323926.
    1. Tilg H, Adolph TE, Gerner RR, Moschen AR. The intestinal microbiota in colorectal cancer. Cancer Cell. 2018;33:954–964. doi: 10.1016/j.ccell.2018.03.004.
    1. Wu S, Rhee KJ, Albesiano E, Rabizadeh S, Wu X, Yen HR, Huso DL, Brancati FL, Wick E, McAllister F, et al. A human colonic commensal promotes colon tumorigenesis via activation of T helper type 17 T cell responses. Nat Med. 2009;15:1016–1022. doi: 10.1038/nm.2015.
    1. Castellarin M, Warren RL, Freeman JD, Dreolini L, Krzywinski M, Strauss J, Barnes R, Watson P, Allen-Vercoe E, Moore RA, Holt RA. Fusobacterium nucleatum infection is prevalent in human colorectal carcinoma. Genome Res. 2012;22:299–306. doi: 10.1101/gr.126516.111.
    1. Arthur JC, Perezchanona E, Mühlbauer M, Tomkovich S, Uronis JM, Fan TJ, Campbell BJ, Abujamel T, Dogan B, Rogers AB, et al. Intestinal inflammation targets cancer-inducing activity of the microbiota. Science. 2012;338:120–123. doi: 10.1126/science.1224820.
    1. Sobhani I, Amiot A, Le Baleur Y, Levy M, Auriault ML, Van Nhieu JT, Delchier JC. Microbial dysbiosis and colon carcinogenesis: Could colon cancer be considered a bacteria-related disease? Therap Adv Gastroenterol. 2013;6:215–229. doi: 10.1177/1756283X12473674.
    1. Isaac A, Kochubiei O, Vizir M. Colorectal cancer treatment. KhNMU. 2014
    1. Xinli L, Dachang W, Cuili Z, Yi X. Side effects of antibiotics on the intestinal microflora by PCR-DGGE. Pak J Pharm Sci. 2013;26:339–343.
    1. Schmidt TM, Delong EF, Pace NR. Analysis of a marine picoplankton community by 16S rRNA gene cloning and sequencing. J Bacteriol. 1991;173:4371–4378. doi: 10.1128/jb.173.14.4371-4378.1991.
    1. Wuyts J, Peer YVD. The European ribosomal RNA database. Nucleic Acids Res. 2004;32:D101–D103. doi: 10.1093/nar/gkh065.
    1. Thompson JR, Marcelino LA, Polz MF. Heteroduplexes in mixed-template amplifications: Formation, consequence and elimination by ‘reconditioning PCR’. Nucleic Acids Res. 2002;30:2083–2088. doi: 10.1093/nar/30.9.2083.
    1. Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, Lesniewski RA, Oakley BB, Parks DH, Robinson CJ, et al. Introducing mothur: Open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol. 2009;75:7537–7541. doi: 10.1128/AEM.01541-09.
    1. Pruesse E, Quast C, Knittel K, Fuchs BM, Ludwig W, Peplies J, Glöckner FO. SILVA: A comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Res. 2007;35:7188–7196. doi: 10.1093/nar/gkm864.
    1. Chao A. Estimating the population size for capture-recapture data with unequal catchability. Biometrics. 1987;43:783–791. doi: 10.2307/2531532.
    1. Chao A, Hwang WH, Chen YC, Kuo CY. Estimating the number of shared species in two communities. Statistica Sinica. 2000;10:227–246.
    1. Shannon CE. The mathematical theory of communication. 1963. MD Comput. 1997;14:306–317.
    1. Simpson EH. Measurement of diversity. Nature. 1949 Apr 30;163 doi: 10.1038/163688a0.
    1. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 1997;25:4876–4882. doi: 10.1093/nar/25.24.4876.
    1. Saitou N, Nei M. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987;4:406–425.
    1. Kumar S, Tamura K, Nei M. MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform. 2004;5:150–163. doi: 10.1093/bib/5.2.150.
    1. Schwabe RF, Jobin C. The microbiome and cancer. Nat Rev Cancer. 2013;13:800–812. doi: 10.1038/nrc3610.
    1. Allen-Vercoe E, Jobin C. Fusobacterium and enterobacteriaceae: Important players for CRC? Immunol Lett. 2014;162:54–61. doi: 10.1016/j.imlet.2014.05.014.
    1. Kim J, Artinyan A, Mailey B, Christopher S, Lee W, McKenzie S, Chen SL, Bhatia S, Pigazzi A, Garcia-Aguilar J. An interaction of race and ethnicity with socioeconomic status in rectal cancer outcomes. Ann Surg. 2011;253:647–654. doi: 10.1097/SLA.0b013e3182111102.
    1. Day GE, Provost E, Lanier AP. Alaska native mortality rates and trends. Public Health Rep. 2009;124:54–64. doi: 10.1177/003335490912400109.
    1. Lanier AP, Day GE, Kelly JJ, Provost E. Disparities in cancer mortality among Alaska Native people, 1994–2003. Alaska Med. 2008;49:120–125.
    1. Yothers G, Sargent DJ, Wolmark N, Goldberg RM, O'Connell MJ, Benedetti JK, Saltz LB, Dignam JJ, Blackstock AW, ACCENT Collaborative Group Outcomes among black patients with stage II and III colon cancer receiving chemotherapy: An analysis of ACCENT adjuvant trials. J Natl Cancer Inst. 2011;103:1498–1506. doi: 10.1093/jnci/djr310.
    1. Hester CM, Jala VR, Langille MG, Umar S, Greiner KA, Haribabu B. Fecal microbes, short chain fatty acids, and colorectal cancer across racial/ethnic groups. World J Gastroenterol. 2015;21:2759–2769. doi: 10.3748/wjg.v21.i9.2759.
    1. Bruno ME, Rogier EW, Frantz AL, Stefka AT, Thompson SN, Kaetzel CS. Regulation of the polymeric immunoglobulin receptor in intestinal epithelial cells by enterobacteriaceae: Implications for mucosal homeostasis. Immunol Invest. 2010;39:356–382. doi: 10.3109/08820131003622809.
    1. Morgan XC, Tickle TL, Sokol H, Gevers D, Devaney KL, Ward DV, Reyes JA, Shah SA, LeLeiko N, Snapper SB, et al. Dysfunction of the intestinal microbiome in inflammatory bowel disease and treatment. Genome Biol. 2012;13:R79. doi: 10.1186/gb-2012-13-9-r79.
    1. Stecher B. The roles of inflammation, nutrient availability and the commensal microbiota in enteric pathogen infection. Microbiol Spectr. 2015;3
    1. Mira-Pascual L, Cabrera-Rubio R, Ocon S, Costales P, Parra A, Suarez A, Moris F, Rodrigo L, Mira A, Collado MC. Microbial mucosal colonic shifts associated with the development of colorectal cancer reveal the presence of different bacterial and archaeal biomarkers. J Gastroenterol. 2015;50:167–179. doi: 10.1007/s00535-014-0963-x.
    1. Garrett WS, Gallini CA, Yatsunenko T, Michaud M, DuBois A, Delaney ML, Punit S, Karlsson M, Bry L, Glickman JN, et al. Enterobacteriaceae act in concert with the gut microbiota to induce spontaneous and maternally transmitted colitis. Cell Host Microbe. 2010;8:292–300. doi: 10.1016/j.chom.2010.08.004.
    1. Lupp C, Robertson ML, Wickham ME, Sekirov I, Champion OL, Gaynor EC, Finlay BB. Host-mediated inflammation disrupts the intestinal microbiota and promotes the overgrowth of enterobacteriaceae. Cell Host Microbe. 2007;2:204. doi: 10.1016/j.chom.2007.06.010.
    1. Holzapfel WH, Wood BJB. The family enterococcaceae. John Wiley Sons Ltd; 2014.
    1. 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. ISME J. 2012;6:320–329. doi: 10.1038/ismej.2011.109.
    1. Balamurugan R, Rajendiran E, George S, Samuel GV, Ramakrishna BS. Real-time polymerase chain reaction quantification of specific butyrate-producing bacteria, Desulfovibrio and Enterococcus faecalis in the feces of patients with colorectal cancer. J Gastroenterol Hepatol. 2008;23:1298–1303. doi: 10.1111/j.1440-1746.2008.05490.x.
    1. Balish E, Warner T. Enterococcus faecalis induces inflammatory bowel disease in interleukin-10 knockout mice. Am J Pathol. 2002;160:2253–2257. doi: 10.1016/S0002-9440(10)61172-8.
    1. Sun T, Liu S, Zhou Y, Yao Z, Zhang D, Cao S, Wei Z, Tan B, Li Y, Lian Z, Wang S. Evolutionary biologic changes of gut microbiota in an ‘adenoma-carcinoma sequence’ mouse colorectal cancer model induced by 1,2-Dimethylhydrazine. Oncotarget. 2017;8:444–457.
    1. Kohoutova D, Smajs D, Moravkova P, Cyrany J, Moravkova M, Forstlova M, Cihak M, Rejchrt S, Bures J. Escherichia coli strains of phylogenetic group B2 and D and bacteriocin production are associated with advanced colorectal neoplasia. BMC Infect Dis. 2014;14:733. doi: 10.1186/s12879-014-0733-7.
    1. Farkas-Himsley H, Yu H. Purified colicin as cytotoxic agent of neoplasia: Comparative study with crude colicin. Cytobios. 1985;42:193–207.
    1. Bonnet M, Buc E, Sauvanet P, Darcha C, Dubois D, Pereira B, Déchelotte P, Bonnet R, Pezet D, Darfeuille-Michaud A. Colonization of the human gut by E. coli and colorectal cancer risk. Clin Cancer Res. 2014;20:859–867. doi: 10.1158/1078-0432.CCR-13-1343.
    1. Peterson CT, Vaughn AR, Sharma V, Chopra D, Mills PJ, Peterson SN, Sivamani RK. Effects of turmeric and curcumin dietary supplementation on human gut microbiota: A double-blind, randomized, placebo-controlled pilot study. J Evid Based Integr Med. 2018;23:2515690X18790725. doi: 10.1177/2515690X18790725.
    1. Stecher B, Robbiani R, Walker AW, Westendorf AM, Barthel M, Kremer M, Chaffron S, Macpherson AJ, Buer J, Parkhill J, et al. Salmonella enterica serovar typhimurium exploits inflammation to compete with the intestinal microbiota. PLos Biol. 2007;5:2177–2189. doi: 10.1371/journal.pbio.0050244.
    1. Gevers D, Kugathasan S, Denson LA, Vázquez-Baeza Y, Van Treuren W, Ren B, Schwager E, Knights D, Song SJ, Yassour M, et al. The treatment-naive microbiome in new-onset Crohn's disease. Cell Host Microbe. 2014;15:382–392. doi: 10.1016/j.chom.2014.02.005.
    1. Wu N, Yang X, Zhang R, Li J, Xiao X, Hu Y, Chen Y, Yang F, Lu N, Wang Z, et al. Dysbiosis signature of fecal microbiota in colorectal cancer patients. Microb Ecol. 2013;66:462–470. doi: 10.1007/s00248-013-0245-9.
    1. Hendrickx AP, Top J, Bayjanov JR, Kemperman H, Rogers MR, Paganelli FL, Bonten MJ, Willems RJ. Antibiotic-driven dysbiosis mediates intraluminal agglutination and alternative segregation of enterococcus faecium from the intestinal epithelium. MBio. 2015;6:e01346–e01315. doi: 10.1128/mBio.01346-15.
    1. Sanchez-Diaz AM, Cuartero C, Rodriguez JD, Lozano S, Alonso JM, Rodríguez-Domínguez M, Tedim AP, Del Campo R, López J, Cantón R, Ruiz-Garbajosa P. The rise of ampicillin-resistant Enterococcus faecium high-risk clones as a frequent intestinal colonizer in oncohaematological neutropenic patients on levofloxacin prophylaxis: A risk for bacteraemia? Clin Microbiol Infect. 2016;22(59):e51–e58.
    1. Becking JH. The family azotobacteraceae. Springer; New York: 2006.
    1. Chen L, Zhang YH, Huang T, Cai YD. Gene expression profiling gut microbiota in different races of humans. Sci Rep. 2016;6:23075. doi: 10.1038/srep23075.
    1. Collins MD, Hutson RA, Foster G, Falsen E, Weiss N. Isobaculum melis gen. nov., sp. nov., a Carnobacterium-like organism isolated from the intestine of a badger. Int J Syst Evol Microbiol. 2002;52:207–210. doi: 10.1099/00207713-52-1-207.
    1. Aller AI, Castro C, Medina MJ, González MT, Sevilla P, Morilla MD, Corzo JE, Martín-Mazuelos E. Isolation of Moellerella wisconsensis from blood culture from a patient with acute cholecystitis. Clin Microbiol Infect. 2009;15:1193–1194. doi: 10.1111/j.1469-0691.2009.03046.x.
    1. Stock I, Falsen E, Wiedemann B. Moellerella wisconsensis: Identification, natural antibiotic susceptibility and its dependency on the medium applied. Diagn Microbiol Infect Dis. 2003;45:1–11. doi: 10.1016/S0732-8893(02)00483-2.
    1. Gagnière J, Raisch J, Veziant J, Barnich N, Bonnet R, Buc E, Bringer MA, Pezet D, Bonnet M. Gut microbiota imbalance and colorectal cancer. World J Gastroenterol. 2016;22:501–518. doi: 10.3748/wjg.v22.i2.501.
    1. Dubrow R, Edberg S, Wikfors E, Callan D, Troncale F, Vender R, Brand M, Yapp R. Fecal carriage of Streptococcus bovis and colorectal adenomas. Gastroenterology. 1991;101:721–725. doi: 10.1016/0016-5085(91)90531-O.
    1. Farhadi T, Fakharian A, Ovchinnikov RS. Virtual screening for potential inhibitors of CTX-M-15 protein of Klebsiella pneumoniae. Interdiscip Sci. 2018;10:694–703. doi: 10.1007/s12539-017-0222-y.
    1. Swidsinski A, Khilkin M, Kerjaschki D, Schreiber S, Ortner M, Weber J, Lochs H. Association between intraepithelial escherichia coli and colorectal cancer. Gastroenterology. 1998;115:281–286. doi: 10.1016/S0016-5085(98)70194-5.
    1. Poulsen ML, Bisgaard ML. MUTYH associated polyposis (MAP) Curr Genomics. 2008;9:420–435. doi: 10.2174/138920208785699562.
    1. Khan AA, Cash P. E. coli and colon cancer: Is mutY a culprit? Cancer Lett. 2013;341:127–131. doi: 10.1016/j.canlet.2013.08.003.
    1. Yu T, Guo F, Yu Y, Sun T, Ma D, Han J, Qian Y, Kryczek I, Sun D, Nagarsheth N, et al. Fusobacterium nucleatum promotes chemoresistance to colorectal cancer by modulating autophagy. Cell. 2017;170(548–563):e16.
    1. Bourassa L, Clarridge JE., III Clinical significance and characterization of streptococcus tigurinus isolates in an adult population. J Clin Microbiol. 2015;53:3574–3579. doi: 10.1128/JCM.01551-15.
    1. Kim B, Huh HJ, Chung DR, Kim WS, Ki CS, Lee NY. The first case of concurrent infective endocarditis and spondylitis caused by streptococcus tigurinus. Ann Lab Med. 2015;35:654–656. doi: 10.3343/alm.2015.35.6.654.
    1. Michelena A, Bonavila C, Zubeltzu B, Goenaga MA. Endocarditis due to Streptococcus tigurinus: Presentation of a case and a review of the literature. Enferm Infecc Microbiol Clín. 2015;33:575–576. doi: 10.1016/j.eimc.2015.01.006. (In Spanish)
    1. Skowron MA, Zebrowska J, Wegrzyn G, Skowron PM. MmoSTI restriction endonuclease, isolated from Morganella morganii infecting a tropical moth, Actias selene, cleaving 5′-|CCNGG-3′ sequences. J Appl Genetics. 2016;57:143–149. doi: 10.1007/s13353-015-0308-3.
    1. Murphy K, Ryan C, Dempsey EM, O'Toole PW, Ross RP, Stanton C, Ryan CA. Neonatal sulfhemoglobinemia and hemolytic anemia associated with intestinal morganella morganii. Pediatrics. 2015;136:1641–1645. doi: 10.1542/peds.2015-0996.
    1. Shahbazi E, Mollasalehi H, Minai-Tehrani D. Development and evaluation of an improved quantitative loop-mediated isothermal amplification method for rapid detection of morganella morganii. Talanta. 2019;191:54–58. doi: 10.1016/j.talanta.2018.08.033.
    1. Koh GY, Kane A, Lee K, Xu Q, Wu X, Roper J, Mason JB, Crott JW. Parabacteroides distasonis attenuates toll-like receptor 4 signaling and Akt activation and blocks colon tumor formation in high-fat diet-fed azoxymethane-treated mice. Int J Cancer. 2018 Apr 26; doi: 10.1002/ijc.31559. (Epub ahead of print)
    1. Dziarski R, Park SY, Kashyap DR, Dowd SE, Gupta D. Pglyrp-regulated gut microflora prevotella falsenii, parabacteroides distasonis and bacteroides eggerthii enhance and alistipes finegoldii attenuates colitis in mice. PLos One. 2016;11:e0146162. doi: 10.1371/journal.pone.0146162.
    1. Davis-Richardson AG, Ardissone AN, Dias R, Simell V, Leonard MT, Kemppainen KM, Drew JC, Schatz D, Atkinson MA, Kolaczkowski B, et al. Bacteroides dorei dominates gut microbiome prior to autoimmunity in Finnish children at high risk for type 1 diabetes. Front Microbiol. 2014;5:678. doi: 10.3389/fmicb.2014.00678.
    1. Ignacio A, Fernandes MR, Avila-Campos MJ, Nakano V. Enterotoxigenic and non-enterotoxigenic bacteroides fragilis from fecal microbiota of children. Braz J Microbiol. 2015;46:1141–1145. doi: 10.1590/S1517-838246420140728.

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