1887

Abstract

In order to characterize the bacterial microbiota present within oral cancerous lesions, tumorous and non-tumorous mucosal tissue specimens (approx. 1 cm) were harvested from ten oral squamous cell carcinoma (OSCC) patients at the time of surgery. Any microbial contamination on the surface of the specimens was eliminated by immersion in Betadine and washing with PBS. Bacteria were visualized within sections of the OSCC by performing fluorescent hybridization with the universal oligonucleotide probe, EUB338. DNA was extracted from each aseptically macerated tissue specimen using a commercial kit. This was then used as template for PCR with three sets of primers, targeting the 16S rRNA genes of , and the domain . PCR products were differentiated by TA cloning and bacterial species were identified by partial sequencing of the 16S rRNA gene fragments. A total of 70 distinct taxa was detected: 52 different phylotypes isolated from the tumorous tissues, and 37 taxa from within the non-tumorous specimens. Differences between the composition of the microbiotas within the tumorous and non-tumorous mucosae were apparent, possibly indicating selective growth of bacteria within carcinoma tissue. Most taxa isolated from within the tumour tissue represented saccharolytic and aciduric species. Whether the presence of these bacteria within the mucosa has any bearing on the carcinogenic process is a concept worthy of further investigation.

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2007-12-01
2024-03-29
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References

  1. Aas J. A., Paster B. J., Stokes L. N., Olsen I., Dewhirst F. E. 2005; Defining the normal bacterial flora of the oral cavity. J Clin Microbiol 43:5721–5732 [CrossRef]
    [Google Scholar]
  2. Agresti A. 1992; A survey of exact inference for contingency tables. Stat Sci 7:131–153 [CrossRef]
    [Google Scholar]
  3. Amann R., Fuchs B. M., Behrens S. 2001; The identification of microorganisms by fluorescence in situ hybridisation. Curr Opin Biotechnol 12:231–236 [CrossRef]
    [Google Scholar]
  4. Baker G. C., Smith J. J., Cowan D. A. 2003; Review and re-analysis of domain-specific 16S primers. J Microbiol Methods 55:541–555 [CrossRef]
    [Google Scholar]
  5. Banerjee A., Yasseri M., Munson M. 2002; A method for the detection and quantification of bacteria in human carious dentine using fluorescent in situ hybridisation. J Dent 30:359–363 [CrossRef]
    [Google Scholar]
  6. Becker M. R., Paster B. J., Leys E. J., Moeschberger M. L., Kenyon S. G., Galvin J. L., Boches S. K., Dewhirst F. E., Griffen A. L. 2002; Molecular analysis of bacterial species associated with childhood caries. J Clin Microbiol 40:1001–1009 [CrossRef]
    [Google Scholar]
  7. Björkholm B., Falk P., Engstrand L., Nyrén O. 2003; Helicobacter pylori : resurrection of the cancer link. J Intern Med 253:102–119 [CrossRef]
    [Google Scholar]
  8. Chen A. Y., Myers J. N. 2001; Cancer of the oral cavity. Dis Mon 47:275–361 [CrossRef]
    [Google Scholar]
  9. Chhour K. L., Nadkarni M. A., Byun R., Martin F. E., Jacques N. A., Hunter N. 2005; Molecular analysis of microbial diversity in advanced caries. J Clin Microbiol 43:843–849 [CrossRef]
    [Google Scholar]
  10. Cole J. R., Chai B., Marsh T. L., Farris R. J., Wang Q., Kulam S. A., Chandra S., McGarrell D. M., Schmidt T. M. other authors 2003; The Ribosomal Database Project (RDP-II): previewing a new autoaligner that allows regular updates and the new prokaryotic taxonomy. Nucleic Acids Res 31:442–443 [CrossRef]
    [Google Scholar]
  11. Coussens L. M., Werb Z. 2002; Inflammation and cancer. Nature 420:860–867 [CrossRef]
    [Google Scholar]
  12. Cross S. S. 2004; Diagnostic pathology in clinical practice. In General and Systematic Pathology , 4th edn. pp 57–70 Edited by Underwood J. C. E. London: Churchill Livingstone;
    [Google Scholar]
  13. Curtis P., Nakatsu C. H., Konopka A. 2002; Aciduric proteobacteria isolated from pH 2.9 soil. Arch Microbiol 178:65–70 [CrossRef]
    [Google Scholar]
  14. Davies C. E., Hill K. E., Wilson M. J., Stephens P., Hill C. M., Harding K. G., Thomas D. W. 2004; Use of 16S ribosomal DNA PCR and denaturing gradient gel electrophoresis for analysis of the microbiotas of healing and nonhealing chronic venous leg ulcers. J Clin Microbiol 42:3549–3557 [CrossRef]
    [Google Scholar]
  15. Dutta U., Garg P. K., Kumar R., Tandon R. K. 2000; Typhoid carriers among patients with gallstones are at increased risk for carcinoma of the gallbladder. Am J Gastroenterol 95:784–787 [CrossRef]
    [Google Scholar]
  16. Ellmerich S., Schöller M., Duranton B., Gossé F., Galluser M., Klein J.-P., Raul F. 2000; Promotion of intestinal carcinogenesis by Streptococcus bovis . Carcinogenesis 21:753–756 [CrossRef]
    [Google Scholar]
  17. Franceschi S., Bidoli E., Herrero R., Muñoz N. 2000; Comparison of cancers of the oral cavity and pharynx worldwide: etiological clues. Oral Oncol 36:106–115 [CrossRef]
    [Google Scholar]
  18. Ha P. K., Califano J. A. 2004; The role of human papillomavirus in oral carcinogenesis. Crit Rev Oral Biol Med 15:188–196 [CrossRef]
    [Google Scholar]
  19. Hindle I., Downer M. C., Moles D. R., Speight P. M. 2000; Is alcohol responsible for more intra-oral cancer?. Oral Oncol 36:328–333 [CrossRef]
    [Google Scholar]
  20. Hooper S. J., Crean S., Lewis M. A. O., Spratt D. A., Wade W. G., Wilson M. J. 2006; Viable bacteria present within oral squamous cell carcinoma tissue. J Clin Microbiol 44:1719–1725 [CrossRef]
    [Google Scholar]
  21. Huber T., Faulkner G., Hugenholtz P. 2004; Bellerophon: a program to detect chimeric sequences in multiple sequence alignments. Bioinformatics 20:2317–2319 [CrossRef]
    [Google Scholar]
  22. Hughes M. S., Beck L.-A., Skuce R. A. 1994; Identification and elimination of DNA sequences in Taq DNA polymerase. J Clin Microbiol 32:2007–2008
    [Google Scholar]
  23. Kazor C. E., Mitchell P. M., Lee A. M., Stokes L. N., Loesche W. J., Dewhirst F. E., Paster B. J. 2003; Diversity of bacterial populations on the tongue dorsa of patients with halitosis and healthy patients. J Clin Microbiol 41:558–563 [CrossRef]
    [Google Scholar]
  24. Knasmüller S., Steinkellner H., Hirschl A. M., Rabot S., Nobis E. C., Kassie F. 2001; Impact of bacteria in dairy products and of the intestinal microbiota on the genotoxic and carcinogenic effects of heterocyclic aromatic amines. Mutat Res 480:481129–138
    [Google Scholar]
  25. Kujan O., Glenny A.-M., Duxbury J., Thakker N., Sloan P. 2005; Evaluation of screening strategies for improving oral cancer mortality: a Cochrane systematic review. J Dent Educ 69:255–265
    [Google Scholar]
  26. Lane D. J. 1991; 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics pp 115–175 Edited by Stackebrandt E., Goodfellow M. Chichester: Wiley;
    [Google Scholar]
  27. Lax A. J. 2005; Bacterial toxins and cancer – case to answer?. Nat Rev Microbiol 3:343–349 [CrossRef]
    [Google Scholar]
  28. Lax A. J., Thomas W. 2002; How bacteria could cause cancer: one step at a time. Trends Microbiol 10:293–299 [CrossRef]
    [Google Scholar]
  29. Lissowska J., Pilarska A., Pilarski P., Samolczyk-Wanyura D., Piekarczyk J., Bardin-Mikolajczak A., Zatonski W., Herrero W., Muñoz N., Franceschi S. 2003; Smoking, alcohol, diet, dentition and sexual practices in the epidemiology of oral cancer in Poland. Eur J Cancer Prev 12:25–33 [CrossRef]
    [Google Scholar]
  30. Llewellyn C. D., Linklater K., Bell J., Johnson N. W., Warnakulasuriya S. 2004; An analysis of risk factors for oral cancer in young people: a case–control study. Oral Oncol 40:304–313 [CrossRef]
    [Google Scholar]
  31. Mager D. L., Haffajee A. D., Devlin P. M., Norris C. M., Posner M. R., Goodson J. M. 2005; The salivary microbiota as a diagnostic indicator of oral cancer: a descriptive, non-randomized study of cancer-free and oral squamous cell carcinoma subjects. J Transl Med 3:27 [CrossRef]
    [Google Scholar]
  32. Marchant S., Brailsford S. R., Twomey A. C., Roberts G. J., Beighton D. 2001; The predominant microbiota of nursing caries lesions. Caries Res 35:397–406 [CrossRef]
    [Google Scholar]
  33. Mohammadi T. H., Reesink W., Vandenbroucke-Grauls C. M. J. E., Savelkoul P. H. M. 2005; Removal of contaminating DNA from commercial nucleic acid extraction kit reagents. J Microbiol Methods 61:285–288 [CrossRef]
    [Google Scholar]
  34. Morita E., Narikiyo M., Yano A., Nishimura E., Igaki H., Sasaki H., Terada M., Hanada N., Kawabe R. 2003; Different frequencies of Streptococcus anginosus infection in oral cancer and esophageal cancer. Cancer Sci 94:492–496 [CrossRef]
    [Google Scholar]
  35. Moter A., Göbel U. B. 2000; Fluorescence in situ hybridization (FISH) for direct visualization of microorganisms. J Microbiol Methods 41:85–112 [CrossRef]
    [Google Scholar]
  36. Munson M. A., Pitt-Ford T., Chong B., Weightman A., Wade W. G. 2002; Molecular and cultural analysis of the microbiota associated with endodontic infections. J Dent Res 81:761–766 [CrossRef]
    [Google Scholar]
  37. Munson M. A., Banerjee A., Watson T. F., Wade W. G. 2004; Molecular analysis of the microbiota associated with dental caries. J Clin Microbiol 42:3023–3029 [CrossRef]
    [Google Scholar]
  38. Nagy K. N., Sonkodi I., Szöde I., Nagy E., Newman H. N. 1998; The microbiota associated with human oral carcinomas. Oral Oncol 34:304–308 [CrossRef]
    [Google Scholar]
  39. Narikiyo M., Tanabe C., Yamada Y., Igaki H., Tachimori Y., Kato H., Muto M., Montesano R., Sakamoto H. other authors 2004; Frequent and preferential infection of Treponema denticola , Streptococcus mitis, and Streptococcus anginosus in esophageal cancers. Cancer Sci 95:569–574 [CrossRef]
    [Google Scholar]
  40. Ogden G. R. 2005; Alcohol and oral cancer. Alcohol 35:169–173 [CrossRef]
    [Google Scholar]
  41. Parsonnet J. 1995; Bacterial infection as a cause of cancer. Environ Health Perspect 103:263–268 [CrossRef]
    [Google Scholar]
  42. Paster B. J., Boches S. K., Galvin J. L., Ericson R. E., Lau C. N., Levanos V. A., Sahasrabudhe A., Dewhirst F. E. 2001; Bacterial diversity in human subgingival plaque. J Bacteriol 183:3770–3783 [CrossRef]
    [Google Scholar]
  43. Paster B. J., Falkner W. A. Jr, Enwonwu C. O., Idigbe E. O., Savage K. O., Levanos V. A., Tamer M. A., Ericson R. L., Lau C. N., Dewhirst F. E. 2002; Prevalent bacterial species and novel phylotypes in advanced noma lesions. J Clin Microbiol 40:2187–2191 [CrossRef]
    [Google Scholar]
  44. Pearson W. R. 1990; Rapid and sensitive sequence comparison with fastp and fasta. Methods Enzymol 183:63–98
    [Google Scholar]
  45. Raghunand N., Gatenby R. A., Gillies R. J. 2003; Microenvironmental and cellular consequences of altered blood flow in tumours. Br J Radiol 76:S11–S22 [CrossRef]
    [Google Scholar]
  46. Reichart P. A. 2001; Identification of risk groups for oral precancer and cancer and preventive measures. Clin Oral Investig 5:207–213 [CrossRef]
    [Google Scholar]
  47. Rosenquist K., Wennerberg J., Schildt E.-B., Bladström A., Hansson B. G., Andersson G. 2005; Oral status, oral infections and some lifestyle factors as risk factors for oral and oropharyngeal squamous cell carcinoma. A population-based case–control study in southern Sweden. Acta Otolaryngol 125:1327–1336 [CrossRef]
    [Google Scholar]
  48. Sakamoto H., Naito H., Ohta Y., Tanakna R., Maeda N., Sasaki J., Nord C. E. 1999; Isolation of bacteria from cervical lymph nodes in patients with oral cancer. Arch Oral Biol 44:789–793 [CrossRef]
    [Google Scholar]
  49. Salaspuro M. P. 2003; Acetaldehyde, microbes, and cancer of the digestive tract. Crit Rev Clin Lab Sci 40:183–208 [CrossRef]
    [Google Scholar]
  50. Sasaki H., Ishizuka T., Muto M., Nezu M., Nakanishi Y., Inagaki Y., Watanabe H., Watanabe H., Terada M. 1998; Presence of Streptococcus anginosus DNA in esophageal cancer, dysplasia of esophagus, and gastric cancer. Cancer Res 58:2991–2995
    [Google Scholar]
  51. Sasaki M., Ohara-Nemoto Y., Tajika S., Kobayashi M., Yamaura C., Kimura S. 2001; Antigenic characterisation of a novel Streptococcus anginosus antigen that induces nitric oxide synthesis by murine peritoneal exudate cells. J Med Microbiol 50:952–958
    [Google Scholar]
  52. Shiga K., Tateda M., Saijo S., Hori T., Sato I., Tateno H., Matsuura K., Takasaka T., Miyagi T. 2001; Presence of Streptococcus infection in extra-oropharyngeal head and neck squamous cell carcinoma and its implication in carcinogenesis. Oncol Rep 8:245–248
    [Google Scholar]
  53. Shukla V. K., Singh H., Pandley M., Upadhyay S. K., Nath G. 2000; Carcinoma of the gallbladder – is it a sequel of typhoid?. Dig Dis Sci 45:900–903 [CrossRef]
    [Google Scholar]
  54. Sitheeque M. A. M., Samaranayake L. P. 2003; Chronic hyperplastic candidosis/candidiasis (candidal leukoplakia). Crit Rev Oral Biol Med 14:253–267 [CrossRef]
    [Google Scholar]
  55. Spratt D. A. 2004; Significance of bacterial identification by molecular biology methods. Endod Topics 9:5–14 [CrossRef]
    [Google Scholar]
  56. Sugano N., Yokoyama K., Oshikawa M., Kumagai K., Takane M., Tanaka H., Ito K. 2003; Detection of Streptococcus anginosus and 8-hydroxy-deoxyguanosine in saliva. J Oral Sci 45:181–184 [CrossRef]
    [Google Scholar]
  57. Sunde P. T., Olsen I., Göbel U. B., Theegarten D., Winter S., Debelian G. J., Tronstad L., Moter A. 2003; Fluorescence in situ hybridization (FISH) for direct visualization of bacteria in periapical lesions of asymptomatic root-filled teeth. Microbiology 149:1095–1102 [CrossRef]
    [Google Scholar]
  58. Svensäter G., Larsson U.-B., Greif E. C. G., Cvitkovitch D. G., Hamilton I. R. 1997; Acid tolerance response and survival by oral bacteria. Oral Microbiol Immunol 12:266–273 [CrossRef]
    [Google Scholar]
  59. Takahashi N. 2003; Acid-neutralizing activity during amino acid fermentation by Porphyromonas gingivalis , Prevotella intermedia and Fusobacterium nucleatum . Oral Microbiol Immunol 18:109–113 [CrossRef]
    [Google Scholar]
  60. Tanner M. A., Goebel B. M., Dojka M. A., Pace N. R. 1998; Specific ribosomal DNA sequences from diverse environmental settings correlate with experimental contaminants. Appl Environ Microbiol 64:3110–3113
    [Google Scholar]
  61. Tateda M., Shiga K., Saijo S., Sone M., Hori T., Yokoyama J., Matsuura K., Takasaka T., Miyagi T. 2000; Streptococcus anginosus in head and neck squamous cell carcinoma: implication in carcinogenesis. Int J Mol Med 6:699–703
    [Google Scholar]
  62. Tezal M., Grossi S. G., Genco R. J. 2005; Is periodontitis associated with oral neoplasms?. J Periodontol 76:406–410 [CrossRef]
    [Google Scholar]
  63. Thurnheer T., Gmur R., Guggenheim B. 2004; Multiplex FISH analysis of a six-species bacterial biofilm. J Microbiol Methods 56:37–47 [CrossRef]
    [Google Scholar]
  64. Vaahtovuo J., Korkeamäki M., Munukka E., Viljanen M. K., Toivanen P. 2005; Quantification of bacteria in human feces using 16S rRNA-hybridization, DNA-staining and flow cytometry. J Microbiol Methods 63:276–286 [CrossRef]
    [Google Scholar]
  65. Velly A. M., Franco E. L., Schlecht N., Pintos J., Kowalski L. P., Oliveira B. V., Curado M. P. 1998; Relationship between dental factors and risk of upper aerodigestive tract cancer. Oral Oncol 34:284–291 [CrossRef]
    [Google Scholar]
  66. Wade W. G. 2004; Non-culturable bacteria in complex commensal populations. Adv Appl Microbiol 54:93–106
    [Google Scholar]
  67. Waisberg J., Matheus C. de O. 2002; Infectious endocarditis from Streptococcus bovis associated with colonic carcinoma: case report and literature review. Arq Gastroenterol 39:177–180 [CrossRef]
    [Google Scholar]
  68. Warnakulasuriya S., Sutherland G., Scully C. 2005; Tobacco, oral cancer, and treatment of dependence. Oral Oncol 41:244–260 [CrossRef]
    [Google Scholar]
  69. Wilson M. J., Weightman A. J., Wade W. G. 1997; Applications of molecular ecology in the characterization of uncultured microorganisms associated with human disease. Rev Med Microbiol 8:91–102 [CrossRef]
    [Google Scholar]
  70. Yu Y. A., Shabahang S., Timiryasova T. M., Zhang Q., Beltz R., Gentschev I., Goebel W., Szalay A. A. 2004; Visualization of tumors and metastases in live animals with bacteria and vaccinia virus encoding light-emitting proteins. Nat Biotechnol 22:313–320 [CrossRef]
    [Google Scholar]
  71. Zakrzewska J. M. 1999; Fortnightly review: oral cancer. BMJ 318:1051–1054 [CrossRef]
    [Google Scholar]
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