Top

BMC Cancer

Published in:

Open Access 01-12-2019 | Streptococci | Research article

The presence of bacteria varies between colorectal adenocarcinomas, precursor lesions and non-malignant tissue

Authors: Caspar Bundgaard-Nielsen, Ulrik T. Baandrup, Lars P. Nielsen, Suzette Sørensen

Published in: BMC Cancer | Issue 1/2019

Abstract

Background

A causal association has been suggested between certain bacteria and colorectal cancer (CRC). Only a few studies have, however, investigated the presence of these bacteria directly in colon tissue with conflicting results. It is thus uncertain which role they may have in prognosis and carcinogenesis of CRC.

Methods

Formalin-fixed and paraffin-embedded (FFPE) colorectal tissue samples from patients diagnosed with colorectal cancer (CRC)(tumor and paired normal tissue, n = 99), adenomas (n = 96), or diverticular disease (n = 104) were tested for the presence and bacterial load of Streptococcus gallolyticus (S. gallolyticus), Fusobacterium nucleatum (F. nucleatum), and Bacteroides fragilis (B. fragilis) using quantitative PCR. A subsequent broader search was conducted on a subset of samples using 16S ribosomal RNA gene sequencing. Finally, to evaluate the prognostic value, the bacterial status was compared to patient outcome.

Results

S. gallolyticus was not detected by qPCR in any of the investigated tissue samples and F. nucleatum and B. fragilis were found to be equally distributed in tumors, paired normal tissue, and diverticula, but significantly less present in adenomas compared to both tumors and diverticula. Neither, F. nucleatum nor B. fragilis status affected the five-year prognosis of the patients. The 16S rRNA gene sequencing data revealed that tumors were associated with the Prevotella genus while conversely adenomas and diverticula were associated with Acinetobacter genus.

Conclusion

These findings do not support a role of F. nucleatum or B. fragilis during colorectal beginning, while S. gallolyticus was not implicated in the colorectal tissue of a Danish population. A potential role of the bacterial genera Prevotella and Acinetobacter was indicated, and requires further investigations.

Available only for authorised users

L a T, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;00:1–22.

Muto T, Bussey H, Morson B. The evolution of cancer of the colon and rectum. Cancer. 1975;36:2251–70.CrossRefPubMed

Volgestein B, Fearon E. A genetic model for colorectal tumorogenesis. Cell. 1990;61:759–67. https://doi.org/10.1016/0092-8674(90)90186-I.CrossRef

Hadac JN, Leystra AA, Paul Olson TJ, Maher ME, Payne SN, Yueh A, et al. Colon tumors with the simultaneous induction of APC, KRAS and PIK3CA mutations still progress through the adenoma-to-carcinoma sequence. Cancer Prev Res (Phila). 2015. https://doi.org/10.1158/1940-6207.CAPR-15-0003.CrossRefPubMed

S a B, Day NE, Luben R, Ferrari P, Slimani N, Norat T, et al. Dietary fibre in food and protection against colorectal cancer in the European prospective investigation into Cancer and nutrition (EPIC): an observational study. Lancet. 2003;361:1496–501.CrossRef

Norat T, Bingham S, Ferrari P, Slimani N, Jenab M, Mazuir M, et al. Meat, fish, and colorectal cancer risk: the European prospective investigation into cancer and nutrition. J Natl Cancer Inst. 2005;97:906–16.CrossRefPubMed

Willett WC, Stampfer MJ, Colditz GA, Rosner BA, Speizer FE. Relation of meat, fat, and fiber intake to the risk of colon cancer in a prospective study among women. N Engl J Med. 1990;323:1664–72.CrossRefPubMed

Nagata C, Shimizu H, Kametani M, Takeyama N, Ohnuma T, Matsushita S. Cigarette smoking, alcohol use, and colorectal adenoma in Japanese men and women. Dis Colon Rectum. 1999;42:337–42.CrossRefPubMed

Pedersen A, Johansen C, Grønbaek M. Relations between amount and type of alcohol and colon and rectal cancer in a Danish population based cohort study. Gut. 2003;52:861–7.CrossRefPubMedPubMedCentral

10.

Etzioni R, Urban N, Ramsey S, McIntosh M, Schwartz S, Reid B, et al. The case for early detection. Nat Rev Cancer. 2003;3:243–52.CrossRefPubMed

11.

Sundhedsstyrelsen. Anbefalinger vedrørende screening for tyk- & endetarmskræft. 2010.

12.

Hewitson P, Glasziou P, Watson E, Towler B, Irwig L. Cochrane systematic review of colorectal cancer screening using the fecal occult blood test (hemoccult): an update. Am J Gastroenterol. 2008;103:1541–9.CrossRefPubMed

13.

WC MC, Mason JM. Enterococcal endocarditis associated with carcinoma of the sigmoid; report of a case. J Med Assoc State Ala. 1951;21:162–6.

14.

Hoppes WL, Lerner PI. Nonenterococcal group-D streptococcal endocarditis caused by Streptococcus bovis. Ann Intern Med. 1974;81:588–93.CrossRefPubMed

15.

Shanan S, Gumaa SA, Sandström G, Abd H. Significant association of Streptococcus bovis with malignant gastrointestinal diseases. Int J Microbiol. 2011;2011:1–5.CrossRef

16.

Gupta a, Madani R, Mukhtar H. Streptococcus bovis endocarditis, a silent sign for colonic tumour. Color Dis. 2010;12:164–71.CrossRef

17.

Corredoira-Sánchez J, García-Garrote F, Rabunal R, López-Roses L, García-País MJ, Castro E, et al. Association between bacteremia due to Streptococcus gallolyticus subsp. gallolyticus (Streptococcus bovis I) and colorectal neoplasia: a case-control study. Clin Infect Dis. 2012;55:491–6.CrossRefPubMed

18.

Sharara AI, Abou Hamdan T, Malli A, El-Halabi MM, Hashash JG, Ghaith OA, et al. Association of Streptococcus bovis endocarditis and advanced colorectal neoplasia: a case-control study. J Dig Dis. 2013;14:382–7. https://doi.org/10.1111/1751-2980.12059. CrossRefPubMed

19.

Abdulamir AS, Hafidh RR, Bakar FA. Molecular detection, quantification, and isolation of Streptococcus gallolyticus bacteria colonizing colorectal tumors: inflammation-driven potential of carcinogenesis via IL-1, COX-2, and IL-8. Mol Cancer. 2010;9:249.CrossRefPubMedPubMedCentral

20.

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:e0119462. https://doi.org/10.1371/journal.pone.0119462.CrossRefPubMedPubMedCentral

21.

Castellarin M, Warren RL, Freeman JD, Dreolini L, Krzywinski M, Strauss J, et al. Fusobacterium nucleatum infection is prevalent in human colorectal carcinoma. Genome Res. 2012;22:299–306.CrossRefPubMedPubMedCentral

22.

Kostic AD, Gevers D, Pedamallu CS, Michaud M, Duke F, Earl AM, et al. Genomic analysis identifies association of fusobacterium with colorectal carcinoma. Genome Res. 2012;22:292–8. https://doi.org/10.1101/gr.126573.111.CrossRefPubMedPubMedCentral

23.

Marchesi JR, Dutilh BE, Hall N, Peters WHM, Roelofs R, Boleij A, et al. Towards the human colorectal cancer microbiome. PLoS One. 2011;6:e20447.CrossRefPubMedPubMedCentral

24.

Bullman S, Pedamallu CS, Sicinska E, Clancy TE, Zhang X, Cai D, et al. Analysis of fusobacterium persistence and antibiotic response in colorectal cancer. Science. 2017;358:1443–8. https://doi.org/10.1126/science.aal5240.CrossRefPubMedPubMedCentral

25.

Mima K, Sukawa Y, Nishihara R, Qian ZR, Yamauchi M, Inamura K, et al. Fusobacterium nucleatum and T cells in colorectal carcinoma. JAMA Oncol. 2015;1:653. https://doi.org/10.1001/jamaoncol.2015.1377.CrossRefPubMedPubMedCentral

26.

Kosumi K, Hamada T, Koh H, Borowsky J, Bullman S, Twombly TS, et al. The amount of Bifidobacterium genus in colorectal carcinoma tissue in relation to tumor characteristics and clinical outcome. Am J Pathol. 2018;48(Suppl 1):S67–9. https://doi.org/10.1016/j.ajpath.2018.08.015.CrossRef

27.

Goodwin AC, Destefano Shields CE, Wu S, Huso DL, Wu X, Murray-Stewart TR, et al. Polyamine catabolism contributes to enterotoxigenic Bacteroides fragilis-induced colon tumorigenesis. Proc Natl Acad Sci U S A. 2011;108:15354–9. https://doi.org/10.1073/pnas.1010203108.CrossRefPubMedPubMedCentral

28.

Ulger Toprak N, Yagci A, Gulluoglu BM, Akin ML, Demirkalem P, Celenk T, et al. A possible role of Bacteroides fragilis enterotoxin in the aetiology of colorectal cancer. Clin Microbiol Infect. 2006;12:782–6. https://doi.org/10.1111/j.1469-0691.2006.01494.x.CrossRefPubMed

29.

Wu S, Rhee K-J, Zhang M, Franco A, Sears CL. Bacteroides fragilis toxin stimulates intestinal epithelial cell shedding and gamma-secretase-dependent E-cadherin cleavage. J Cell Sci. 2007;120(Pt 11):1944–52. https://doi.org/10.1242/jcs.03455.CrossRefPubMed

30.

Wu S, Morin PJ, Maouyo D, Sears CL. Bacteroides fragilis enterotoxin induces c-Myc expression and cellular proliferation. Gastroenterology. 2003;124:392–400.CrossRefPubMed

31.

Wei Z, Cao S, Liu S, Yao Z, Sun T, Li Y, et al. Could gut microbiota serve as prognostic biomarker associated with colorectal cancer patients’ survival? A pilot study on relevant mechanism. Oncotarget. 2016;7(29):46158–72. https://doi.org/10.18632/oncotarget.10064.CrossRefPubMedPubMedCentral

32.

Kostic AD, Chun E, Robertson L, Glickman JN, Gallini CA, Michaud M, et al. Fusobacterium nucleatum potentiates intestinal tumorigenesis and modulates the tumor-immune microenvironment. Cell Host Microbe. 2013;14:207–15. https://doi.org/10.1016/j.chom.2013.07.007.CrossRefPubMedPubMedCentral

33.

Zeller G, Tap J, Voigt AY, Sunagawa S, Kultima JR, Costea PI, et al. Potential of fecal microbiota for early-stage detection of colorectal cancer. Mol Syst Biol. 2014;10:766. https://doi.org/10.15252/msb.20145645.CrossRefPubMedPubMedCentral

34.

Rezasoltani S, Asadzadeh Aghdaei H, Dabiri H, Akhavan Sepahi A, Modarressi MH, Nazemalhosseini Mojarad E. The association between fecal microbiota and different types of colorectal polyp as precursors of colorectal cancer. Microb Pathog. 2018;124:244–9. https://doi.org/10.1016/j.micpath.2018.08.035.CrossRefPubMed

35.

McCoy AN, Araújo-Pérez F, Azcárate-Peril A, Yeh JJ, Sandler RS, Keku TO. Fusobacterium is associated with colorectal adenomas. PLoS One. 2013;8:e53653. https://doi.org/10.1371/journal.pone.0053653.CrossRefPubMedPubMedCentral

36.

Park CH, Han DS, Oh YH, Lee AR, Lee YR, Eun CS. Role of fusobacteria in the serrated pathway of colorectal carcinogenesis. Sci Rep. 2016;6:1–8. https://doi.org/10.1038/srep25271.CrossRef

37.

Pagnini C, Corleto VD, Mangoni ML, Pilozzi E, Torre MS, Marchese R, et al. Alteration of local microflora and α-defensins hyper-production in colonic adenoma mucosa. J Clin Gastroenterol. 2011;45:602–10.CrossRefPubMed

38.

Shen XJ, Rawls JF, Randall T, Burcal L, Mpande CN, Jenkins N, et al. Molecular characterization of mucosal adherent bacteria and associations with colorectal adenomas. Gut Microbes. 2010;1:138–47. https://doi.org/10.4161/gmic.1.3.12360.CrossRefPubMedPubMedCentral

39.

Yu J, Chen Y, Fu X, Zhou X, Peng Y, Shi L, et al. Invasive fusobacterium nucleatum may play a role in the carcinogenesis of proximal colon cancer through the serrated neoplasia pathway. Int J Cancer. 2016;139:1318–26.CrossRefPubMed

40.

Feng Q, Liang S, Jia H, Stadlmayr A, Tang L, Lan Z, et al. Gut microbiome development along the colorectal adenoma-carcinoma sequence. Nat Commun. 2015;6:6528.CrossRefPubMed

41.

Yu J, Feng Q, Wong SH, Zhang D, Liang QY, Qin Y, et al. Metagenomic analysis of faecal microbiome as a tool towards targeted non-invasive biomarkers for colorectal cancer. Gut. 2017;66:70–8. https://doi.org/10.1136/gutjnl-2015-309800.CrossRefPubMed

42.

Nakatsu G, Li X, Zhou H, Sheng J, Wong SH, Wu WKK, et al. Gut mucosal microbiome across stages of colorectal carcinogenesis. Nat Commun. 2015;6:8727. https://doi.org/10.1038/ncomms9727.CrossRefPubMed

43.

Gevers D, Kugathasan S, Denson LA, Vázquez-Baeza Y, Van Treuren W, Ren B, et al. The treatment-naive microbiome in new-onset Crohn’s disease. Cell Host Microbe. 2014;15:382–92. https://doi.org/10.1016/j.chom.2014.02.005.CrossRefPubMedPubMedCentral

44.

Promeza. Compare Two Prevalences: Sample Size. http://www.promesa.co.nz/help/EP_ss_compare_prev.htm. Accessed 1 Sept 2015.

45.

Ye J, Coulouris G, Zaretskaya I, Cutcutache I, Rozen S, Madden TL. Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics. 2012;13:134.CrossRefPubMedPubMedCentral

46.

Karlsen F, Kalantari M, Chitemerere M, Johansson B, Hagmar B. Modifications of human and viral deoxyribonucleic acid by formaldehyde fixation. Lab Investig. 1994;71:604–11 http://www.ncbi.nlm.nih.gov/pubmed/7967515.

47.

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^(−ΔΔCT) method. Methods. 2001;25:402–8.CrossRefPubMed

48.

Saghaei M. An overview of randomization and minimization programs for randomized clinical trials. J Med Signals Sens. 2011;1:55–61 http://www.ncbi.nlm.nih.gov/pubmed/22606659. Accessed 13 Oct 2017.PubMedPubMedCentral

49.

Illumina. 16S Metagenomic Sequencing Library Preparation, Part # 15044223 Rev. B. 2015;:1–28. http://support.illumina.com/content/dam/illumina-support/documents/documentation/chemistry_documentation/16s/16s-metagenomic-library-prep-guide-15044223-b.pdf. Accessed 10 Feb 2018.

50.

Albertsen M, Karst SM, Ziegler AS, Kirkegaard RH, Nielsen PH. Back to basics - the influence of DNA extraction and primer choice on phylogenetic analysis of activated sludge communities. PLoS One. 2015;10:1–15.CrossRef

51.

Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Lozupone CA, Turnbaugh PJ, et al. Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample. Proc Natl Acad Sci. 2011;108(Supplement_1):4516–22. https://doi.org/10.1073/pnas.1000080107.CrossRefPubMed

52.

Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Huntley J, Fierer N, et al. Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms. ISME J. 2012;6:1621–4. https://doi.org/10.1038/ismej.2012.8.CrossRefPubMedPubMedCentral

53.

Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014;30:2114–20. https://doi.org/10.1093/bioinformatics/btu170.CrossRefPubMedPubMedCentral

54.

Edgar RC. UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nat Methods. 2013;10:996–8. https://doi.org/10.1038/nmeth.2604.CrossRefPubMed

55.

Wang Q, Garrity GM, Tiedje JM, Cole JR. Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol. 2007;73:5261–7. https://doi.org/10.1128/AEM.00062-07.CrossRefPubMedPubMedCentral

56.

Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, et al. QIIME allows analysis of high-throughput community sequencing data. Nat Methods. 2010;7:335–6. https://doi.org/10.1038/nmeth.f.303.CrossRefPubMedPubMedCentral

57.

McIlroy SJ, Saunders AM, Albertsen M, Nierychlo M, McIlroy B, Hansen AA, et al. MiDAS: the field guide to the microbes of activated sludge. Database. 2015;2015:1–8.CrossRef

58.

R Development Core Team. R: A Language and Enviroment for Statistical Computing. the R Foundation for Statistical Computing. https://www.r-project.org/. Accessed 1 July 2018.

59.

Andersen KSS, Kirkegaard RH, Karst SM, Albertsen M. ampvis2: an R package to analyse and visualise 16S rRNA amplicon data. bioRxiv. 2018:299537. https://doi.org/10.1101/299537.

60.

Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014;15:1–21.CrossRef

61.

Benjamini Y, Yosef H. Determination of water content in meat pâté by karl fischer titration and its moisture sorption characteristics. J R Stat Soc Ser B. 1995;57:289–300.

62.

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;74:1311–8.CrossRefPubMedPubMedCentral

63.

Wu S, Rhee K-J, Albesiano E, Rabizadeh S, Wu X, Yen H-R, et al. A human colonic commensal promotes colon tumorigenesis via activation of T helper type 17 T cell responses. Nat Med. 2009;15:1016–22. https://doi.org/10.1038/nm.2015.CrossRefPubMedPubMedCentral

64.

Arthur JC, Perez-Chanona E, Mühlbauer M, Tomkovich S, Uronis JM, Fan T, et al. Intestinal inflammation targets cancer-inducing activity of the microbiota. Science. 2012;338:120–3. https://doi.org/10.1126/science.1224820.CrossRefPubMedPubMedCentral

65.

Brenner H, Hoffmeister M, Stegmaier C, Brenner G, Altenhofen L, Haug U. Risk of progression of advanced adenomas to colorectal cancer by age and sex: estimates based on 840 149 screening colonoscopies. Gut. 2007;56:1585–9.CrossRefPubMedPubMedCentral

66.

Sobhani I, Tap J, Roudot-Thoraval F, Roperch JP, Letulle S, Langella P, et al. Microbial dysbiosis in colorectal cancer (CRC) patients. PLoS One. 2011;6:e16393.CrossRefPubMedPubMedCentral

67.

Flemer B, Lynch DB, Brown JMR, Jeffery IB, Ryan FJ, Claesson MJ, et al. Tumour-associated and non-tumour-associated microbiota in colorectal cancer. Gut. 2017;66:633–43.CrossRefPubMed

68.

Rubinstein MR, Wang X, Liu W, Hao Y, Cai G, Han YW. Fusobacterium nucleatum promotes colorectal carcinogenesis by modulating E-cadherin/β-catenin signaling via its FadA Adhesin. Cell Host Microbe. 2013;14:195–206. https://doi.org/10.1016/j.chom.2013.07.012.CrossRefPubMedPubMedCentral

69.

Larsen JM. The immune response to Prevotella bacteria in chronic inflammatory disease. Immunology. 2017;151:363–74. https://doi.org/10.1111/imm.12760.CrossRefPubMedPubMedCentral

70.

Thomas AM, Jesus EC, Lopes A, Aguiar S, Begnami MD, Rocha RM, et al. Tissue-associated bacterial alterations in rectal carcinoma patients revealed by 16S rRNA community profiling. Front Cell Infect Microbiol. 2016:1–13. https://doi.org/10.3389/fcimb.2016.00179.

71.

Boleij A, Van Gelder MMHJ, Swinkels DW, Tjalsma H. Clinical importance of streptococcus gallolyticus infection among colorectal cancer patients: systematic review and meta-analysis. Clin Infect Dis. 2011;53:870–8.CrossRefPubMed

72.

Imrit K, Goldfischer M, Wang J, Green J, Levine J, Lombardo J, et al. Identification of bacteria in formalin-fixed, paraffin-embedded heart valve tissue via 16S rRNA gene nucleotide sequencing. J Clin Microbiol. 2006;44:2609–11.CrossRefPubMedPubMedCentral

73.

Bufill JA. Colorectal cancer: evidence for distinct genetic categories based on proximal or distal tumor location. Ann Intern Med. 1990;113:779–88 http://www.ncbi.nlm.nih.gov/pubmed/2240880.CrossRefPubMed

74.

International Union against Cancer (UICC). Sobin LH, Gospodarowicz MK, Wittekind C, editors. TNM : classification of malignant tumours. 7th ed. Chichester: Wiley-Blackwell; 2010.

Title: The presence of bacteria varies between colorectal adenocarcinomas, precursor lesions and non-malignant tissue
Authors: Caspar Bundgaard-Nielsen
Ulrik T. Baandrup
Lars P. Nielsen
Suzette Sørensen
Publication date: 01-12-2019
Publisher: BioMed Central
Keywords: Streptococci
Respiratory Microbiota
Colorectal Cancer
Colorectal Cancer
Published in: BMC Cancer / Issue 1/2019
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/s12885-019-5571-y

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2019

Can ELABELA be a novel target in the treatment of chronic lymphocytic leukaemia?

The small molecule Bcl-2/Mcl-1 inhibitor TW-37 shows single-agent cytotoxicity in neuroblastoma cell lines

Adjuvant cytokine-induced killer cell immunotherapy for hepatocellular carcinoma: a propensity score-matched analysis of real-world data

Clinical and prognostic implications of pretreatment albumin to C-reactive protein ratio in patients with hepatocellular carcinoma

Best fitting tumor growth models of the von Bertalanffy-PütterType

Acquired immunodeficiency associated with thymoma: a case report

Keynote webinar | Spotlight on antibody–drug conjugates in cancer