Chicken or the Egg: Microbial Alterations in Biopsy Samples of Patients with Oral Potentially Malignant Disorders

  • Gabor Decsi
  • Jozsef Soki
  • Bernadett Pap
  • Gabriella Dobra
  • Maria Harmati
  • Sandor Kormondi
  • Tibor Pankotai
  • Gabor Braunitzer
  • Janos Minarovits
  • Istvan Sonkodi
  • Edit Urban
  • Istvan Balazs Nemeth
  • Katalin Nagy
  • Krisztina Buzas
Original Article


Oral carcinogenesis often leads to the alteration of the microbiota at the site of the tumor, but data are scarce regarding the microbial communities of oral potentially malignant disorders (OPMDs). Punch biopsies were taken from healthy and non-healthy mucosa of OPMD patients to analyze the microbiome using metagenome sequencing. In healthy oral mucosa biopsies the bacterial phyla Firmicutes, Fusobacteria, Proteobacteria, Actinobacteria and Bacteroidetes were detected by Ion Torrent sequencing. The same phyla as well as the phyla Fibrobacteres and Spirochaetes were present in the OPMD biopsies. On the species level, there were 10 bacterial species unique to the healthy tissue and 35 species unique to the OPMD lesions whereas eight species were detected in both samples. We observed that the relative abundance of Streptococcus mitis decreased in the OPMD lesions compared to the uninvolved tissue. In contrast, the relative abundance of Fusobacterium nucleatum, implicated in carcinogenesis, was elevated in OPMD. We detected markedly increased bacterial diversity in the OPMD lesions compared to the healthy oral mucosa. The ratio of S. mitis and F. nucleatum are characteristically altered in the OPMD lesions compared to the healthy mucosa.


OPMD Oral microbiome Metagenome sequencing Lichen Leukoplakia 



This work was in part supported by the National Research, Development and Innovation Fund of Hungary, financed under the NKFI-6-K funding scheme (11493 project), GINOP-2.3.2-15-2016-00015, János Bolyai Research Scholarship of the Hungarian Academy of Sciences and GINOP 2.3.2-15-2016-00011.

Compliance with Ethical Standards

Conflict of Interest

No potential conflict of interest was reported by the authors.

Ethical Approval

The study protocol conformed to the Declaration of Helsinki in all respects and was approved by the Institutional Ethics Committee of the University of Szeged (No. 3161).


  1. 1.
    Morse DE, Psoter WJ, Cleveland D et al (2007) Smoking and drinking in relation to oral cancer and oral epithelial dysplasia. Cancer Causes Control 18(9):919–929CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Jepsen SA, Closmann JJ (2008) The insidious nature and presentation of oral squamous cell carcinoma in the low-risk population. Gen Dent 56(1):78–82PubMedGoogle Scholar
  3. 3.
    Llewellyn CD, Linklater K, Bell J et al (2003) Squamous cell carcinoma of the oral cavity in patients aged 45 years and under: a descriptive analysis of 116 cases diagnosed in the South East of England from 1990 to 1997. Oral Oncol 39(2):106–114CrossRefPubMedGoogle Scholar
  4. 4.
    Nagy KN, Sonkodi I, Szöke I et al (1998) The microflora associated with human oral carcinomas. Oral Oncol 34(4):304–308CrossRefPubMedGoogle Scholar
  5. 5.
    Sarode SC, Sarode GS, Tupkari JV (2012) Oral potentially malignant disorders: precising the definition. Oral Oncol 48(9):759–760CrossRefPubMedGoogle Scholar
  6. 6.
    van der Waal I (2009) Potentially malignant disorders of the oral and oropharyngeal mucosa; terminology, classification and present concepts of management. Oral Oncol 45(4–5):317–323CrossRefPubMedGoogle Scholar
  7. 7.
    Sonkodi I (2009) Oral and Maxilofacial medicine. Semmelweis Publisher, BudapestGoogle Scholar
  8. 8.
    Silverman S Jr, Gorsky M (1997) Proliferative verrucous leukoplakia. A follow-up study of 54 cases. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 84(2):154–157CrossRefPubMedGoogle Scholar
  9. 9.
    Fehér E, Gáll T, Murvai M et al (2009) Investigation of the occurrence of torque tenovirus in malignant and potentially malignant disorders associated with human papillomavirus. J Med Virol 81(11):1975–1981CrossRefPubMedGoogle Scholar
  10. 10.
    Fehér E, Kardos G, Gáll T et al (2011) Comparison of diversity of torque teno virus 1 in different mucosal tissues and disorders. Acta Microbiol Immunol Hung 58(4):319–337CrossRefPubMedGoogle Scholar
  11. 11.
    Kis A, Fehér E, Gáll T et al (2009) Epstein-Barr virus prevalence in oral squamous cell cancer and in potentially malignant oral disorders in an eastern Hungarian population. Eur J Oral Sci 117(5):536–540CrossRefPubMedGoogle Scholar
  12. 12.
    Wu L, Feng J, Shi L et al (2013) Candidal infection in oral leukoplakia: a clinicopathologic study of 396 patients from eastern China. Ann Diagn Pathol 17(1):37–40CrossRefPubMedGoogle Scholar
  13. 13.
    Hebbar PB, Pai A, D S. (2013) Mycological and histological associations of Candida in oral mucosal lesions. J Oral Sci 55(2):157–160CrossRefPubMedGoogle Scholar
  14. 14.
    Dilhari A, Weerasekera MM, Siriwardhana A et al (2016) Candida infection in oral leukoplakia: an unperceived public health problem. Acta Odontol Scand 74(7):565–569CrossRefPubMedGoogle Scholar
  15. 15.
    Gainza-Cirauqui ML, Nieminen MT, Novak Frazer L et al (2013) Production of carcinogenic acetaldehyde by Candida albicans from patients with potentially malignant oral mucosal disorders. J Oral Pathol Med. 42(3):243–249CrossRefPubMedGoogle Scholar
  16. 16.
    Bakri MM, Cannon RD, Holmes AR et al (2014) Detection of Candida albicans ADH1 and ADH2 mRNAs in human archival oral biopsy samples. J Oral Pathol Med 43(9):704–710CrossRefPubMedGoogle Scholar
  17. 17.
    Kazanowska-Dygdała M, Duś I, Radwan-Oczko M (2016) The presence of helicobacter pylori in oral cavities of patientsn with leukoplakia and oral lichenplanus. J Appl Oral Sci 24(1):18–23CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Mizuki H, Kawamura T, Nagasawa D (2015) In situ immunohistochemical detection of intracellular mycoplasma salivarium in the epithelial cells of oral leukoplakia. J Oral Pathol Med. 44(2):134–144CrossRefPubMedGoogle Scholar
  19. 19.
    Meisel P, Holtfreter B, Biffar R et al (2012) Association of periodontitis with the risk of oral leukoplakia. Oral Oncol 48(9):859–863CrossRefPubMedGoogle Scholar
  20. 20.
    Hu X, Zhang Q, Hua H et al (2016) Changes in the salivary microbiota of oral leukoplakia and oral cancer. Oral Oncol 56:e6–e8CrossRefPubMedGoogle Scholar
  21. 21.
    Hernandez BY, Zhu X, Goodman MT et al (2017) Betel nut chewing, oral premalignant lesions, and the oral microbiome. PLoS One 12(2):e0172196CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Srinivas K, Aravinda K, Ratnakar P et al (2011) Oral lichen planus – Review on etiopathogenesis. Natl J Maxillofac Surg 2(1):15–16CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Payeras MR, Cherubini K, Figueiredo MA et al (2013) Oral lichen planus: focus on etiopathogenesis. Arch Oral Biol 58(9):1057–1069CrossRefPubMedGoogle Scholar
  24. 24.
    Georgakopoulou EA, Achtari MD, Achtaris M et al (2012) Oral lichen planus as a preneoplastic inflammatory model. J Biomed Biotechnol 2012:759626CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Roopashree MR, Gondhalekar RV, Shashikanth MC et al (2010) Pathogenesis of oral lichen planus--a review. J Oral Pathol Med. 39(10):729–734CrossRefPubMedGoogle Scholar
  26. 26.
    Gorouhi F, Davari P, Fazel N (2014) Cutaneous and mucosal lichen planus: a comprehensive review of clinical subtypes, risk factors, diagnosis, and prognosis. ScientificWorld Journal 2014:742826CrossRefPubMedGoogle Scholar
  27. 27.
    Bäckman K, Jontell M (2007) Microbial-associated oral lichenoid reactions. Oral Dis 13(4):402–406CrossRefPubMedGoogle Scholar
  28. 28.
    Taghavi Zenouz A, Mehdipour M, Jafari Heydarlou M et al (2010) Relationship between Lichen Planus and Helicobacter pylori Infection. J Dent Res Dent Clin Dent Prospects 4(1):17–20PubMedPubMedCentralGoogle Scholar
  29. 29.
    Attia EA, Abdel Fattah NS, Abdella HM (2010) Upper gastrointestinal findings and detection of helicobacter pylori in patients with oral lichen planus. Clin Exp Dermatol 35(4):355–360CrossRefPubMedGoogle Scholar
  30. 30.
    Pourshahidi S, Fakhri F, Ebrahimi H et al (2012) Lack of association between helicobacter pylori infection and oral lichen planus. Asian Pac J Cancer Prev 13(5):1745–1747CrossRefPubMedGoogle Scholar
  31. 31.
    Izol B, Karabulut AA, Biyikoglu I et al (2010) Investigation of upper gastrointestinal tract involvement and H. Pylori presence in lichen planus: a case-controlled study with endoscopic and histopathological findings. Int J Dermatol 49(10):1121–1126CrossRefPubMedGoogle Scholar
  32. 32.
    Hulimavu SR, Mohanty L, Tondikulam NV et al (2014) No evidence for Helicobacter pylori in oral lichen planus. J Oral Pathol Med. 43(8):576–578CrossRefPubMedGoogle Scholar
  33. 33.
    Boorghani M, Gholizadeh N, Taghavi Zenouz A et al (2010) Oral lichen planus: clinical features, etiology, treatment and management; a review of literature. J Dent Res Dent Clin Dent Prospects. 4(1):3–9PubMedPubMedCentralGoogle Scholar
  34. 34.
    Ertugrul AS, Arslan U, Dursun R et al (2013) Periodontopathogen profile of healthy and oral lichen planus patients with gingivitis or periodontitis. Int J Oral Sci 5(2):92–97CrossRefPubMedGoogle Scholar
  35. 35.
    El-Naggar AK, Chan JKC, Takata T, Grandis JR, Slootweg PJ (2017) Tumours of the oral cavity and mobile tongue In: The fourth edition of the head and neck World Health Organization blue book: editors' perspectives. Hum Pathol 66:10–12CrossRefPubMedGoogle Scholar
  36. 36.
    Sharma PK, Capalash N, Kaur J (2007) An improved method for single step purification of metagenomic DNA. Mol Biotechnol 36(1):61–63CrossRefPubMedGoogle Scholar
  37. 37.
    Periasamy S, Chalmers NI, Du-Thumm L et al (2009) Fusobacterium nucleatum ATCC 10953 requires Actinomyces naeslundii ATCC 43146 for growth on saliva in a three-species community that includes Streptococcus oralis 34. Appl Environ Microbiol 75(10):3250–3257CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Rubinstein MR, Wang X, Liu W et al (2013) Fusobacterium nucleatum promotes colorectal carcinogenesis by modulating E-cadherin/β-catenin signaling via its FadA adhesion. Cell Host Microbe 14(2):195–206CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Schmidt BL, Kuczynski J, Bhattacharya A et al (2014) Changes in abundance of oral microbiota associated with oral cancer. PLoS One 9(6):e98741CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Al-Hebshi NN, Nasher AT, Maryoud MY et al (2017) Inflammatory bacteriome featuring fusobacterium nucleatum and Pseudomonas aeruginosa identified in association with oral squamous cell carcinoma. Sci Rep 7(1):1834CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Arduino PG, Romano F, Sasia D et al (2017) Subgingival microbiota in white patients with Desquamative gingivitis: a cross-sectional study. J Periodontol 88(7):643–650CrossRefPubMedGoogle Scholar
  42. 42.
    Kreth J, Merritt J, Qi F (2009) Bacterial and host interactions of oral streptococci. DNA Cell Biol 28(8):397–403CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Van Hoogmoed CG, Geertsema-Doornbusch GI, Teughels W et al (2008) Reduction of periodontal pathogens adhesion by antagonistic strains. Oral Microbial Immunol 23(1):43–48CrossRefGoogle Scholar
  44. 44.
    Standar K, Kreikemeyer B, Redanz S et al (2010) Setup of an in vitro test system for basic studies on biofilm behavior of mixed-species cultures with dental and periodontal pathogens. PLoS One 5(10):e13135CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Otsuki K, Imai N (2017) Effects of lactoferrin in 6 patients with refractory bacterial vaginosis. Biochem Cell Biol 95(1):31–33CrossRefPubMedGoogle Scholar
  46. 46.
    Klarić M, Mandić V, Lovrić S, e al. (2017) Antimicrobial efficacy of probiotic-containing toothpastes: an in vitro evaluation. Med Glas (Zenica) 14(1):16–24Google Scholar

Copyright information

© Arányi Lajos Foundation 2018

Authors and Affiliations

  • Gabor Decsi
    • 1
  • Jozsef Soki
    • 2
  • Bernadett Pap
    • 3
  • Gabriella Dobra
    • 3
  • Maria Harmati
    • 3
  • Sandor Kormondi
    • 4
  • Tibor Pankotai
    • 5
  • Gabor Braunitzer
    • 6
  • Janos Minarovits
    • 7
  • Istvan Sonkodi
    • 1
  • Edit Urban
    • 2
  • Istvan Balazs Nemeth
    • 8
  • Katalin Nagy
    • 1
  • Krisztina Buzas
    • 3
    • 7
  1. 1.Faculty of Dentistry, Department of Oral SurgeryUniversity of SzegedSzegedHungary
  2. 2.Albert Szent-Gyorgyi Clinical Centre, Institute of Clinical MicrobiologyUniversity of SzegedSzegedHungary
  3. 3.Biological Research CentreHungarian Academy of SciencesSzegedHungary
  4. 4.Albert Szent-Gyorgyi Clinical Centre, Department of TraumatologyUniversity of SzegedSzegedHungary
  5. 5.Department of Biochemistry and Molecular BiologyUniversity of SzegedSzegedHungary
  6. 6.dicomLAB Ltd.SzegedHungary
  7. 7.Faculty of Dentistry, Department of Oral Biology and Experimental Dental ResearchUniversity of SzegedSzegedHungary
  8. 8.Department of Dermatology and AllergologyUniversity of SzegedSzegedHungary

Personalised recommendations