Skip to main content

Advertisement

SpringerLink
Log in

Mass Spectrometry-Based Salivary Proteomics for the Discovery of Head and Neck Squamous Cell Carcinoma

  • Research
  • Published:
Pathology & Oncology Research

Abstract

The 5-year survival rates for cases of head and neck squamous cell carcinoma (HNSCC) are only some 60%, mainly because 20%–40% of the patients develop a local relapse in the same or an adjacent anatomic region, even when the surgical margins are histologically tumour-free. Tumours are often discovered in an advanced stage because of the lack of specific symptoms and the diagnostic difficulties. The more advanced the stage of the tumour, the more invasive the diagnostic and treatment interventions needed. An early molecular diagnosis is therefore of vital importance in order to increase the survival rate. The aim of this study was to develop an efficient rapid and sensitive mass spectrometric method for the detection of differentially expressed proteins as tumour-specific biomarkers in saliva from HNSCC patients. Whole saliva samples were collected from patients with HNSCC and from healthy subjects. The proteins were profiled by using SDS PAGE, MALDI TOF/TOF mass spectrometry and the Mascot database search engine. Several potential tumour markers were identified, including annexin A1, beta- and gamma-actin, cytokeratin 4 and 13, zinc finger proteins and P53 pathway proteins. All of these proteins play a proven role in tumour genesis, and have not been detected previously in saliva. Salivary proteomics is a non-invasive specific method for cancer diagnosis and follow-up treatment. It provides facilities for the readily reproducible and reliable detection of tumours in early stages.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Parkin DM, Bray F, Ferlay J, Pisani P (2005) Global cancer statistics, 2002. CA Cancer J Clin 55:74–108

    Article  PubMed  Google Scholar 

  2. Argiris A, Karamouzis MV, Raben D et al (2008) Head and neck cancer. Lancet 371:1695–1709

    Article  PubMed  CAS  Google Scholar 

  3. Mackenzie J, Ah-See K, Thakker N, Sloan P, Maran AG, Birch J et al (2000) Increasing incidence of oral cancer amongst young persons: what is the aetiology? Oral Oncol 36:387–389

    Article  PubMed  CAS  Google Scholar 

  4. Blot WJ, McLaughlin JK, Winn DM, Austin DF, Greenberg RS, Preston-Martin S et al (1988) Smoking and drinking in relation to oral and pharyngeal cancer. Cancer Res 48:3282–3287

    PubMed  CAS  Google Scholar 

  5. Ide R, Mizoue T, Fujino Y, Hoshiyama Y, Sakata K, Tamakoshi A et al (2008) Cigarette smoking, alcohol drinking, and oral and pharyngeal cancer mortality in Japan. Oral Dis 14:314–319

    Article  PubMed  CAS  Google Scholar 

  6. Decker J, Goldstein JC (1982) Risk factors in head and neck cancer. N Engl J Med 306:1151–1155

    Article  PubMed  CAS  Google Scholar 

  7. Hashibe M, Brennan P, Benhamou S, Castellsague X, Chen C, Curado MP et al (2007) Alcohol drinking in never users of tobacco, cigarette smoking in never drinkers, and the risk of head and neck cancer: pooled analysis in the International Head and Neck Cancer Epidemiology Consortium. J Natl Cancer Inst 99:777–789

    Article  PubMed  Google Scholar 

  8. D’Souza G, Kreimer AR, Viscidi R, Pawlita M, Fakhry C, Koch WM et al (2007) Case–control study of human papillomavirus and oropharyngeal cancer. N Engl J Med 356:1944–1956

    Article  PubMed  Google Scholar 

  9. Wu L et al (2000) Evidence for a causal association between human papillomavirus and a subset of head and neck cancers. J Natl Cancer Inst 92:709–720

    Article  PubMed  Google Scholar 

  10. Gillison ML, Koch WM, Shah KV (1999) Human papillomavirus in head and neck squamous cell carcinoma: are some head and neck cancers a sexually transmitted disease? Curr Opin Oncol 11:191–199

    Article  PubMed  CAS  Google Scholar 

  11. Gillison ML, Lowy DR (2004) A causal role for human papillomavirus in head and neck cancer. Lancet 363:1488–1489

    Article  PubMed  CAS  Google Scholar 

  12. Kreimer AR, Alberg AJ, Daniel R, Gravitt PE, Viscidi R, Garrett ES et al (2004) Oral human papillomavirus infection in adults is associated with sexual behavior and HIV serostatus. J Infect Dis 189:686–698

    Article  PubMed  Google Scholar 

  13. Kreimer AR, Clifford GM, Boyle P, Franceschi S (2005) Human papillomavirus types in head and neck squamous cell carcinomas worldwide: a systematic review. Cancer Epidemiol Biomarkers Prev 14:467–475

    Article  PubMed  CAS  Google Scholar 

  14. Mork J, Lie AK, Glattre E, Hallmans G, Jellum E, Koskela P et al (2001) Human papillomavirus infection as a risk factor for squamous-cell carcinoma of the head and neck. N Engl J Med 344:1125–1131

    Article  PubMed  CAS  Google Scholar 

  15. Mendenhall WM, Logan HL (2009) Human papillomavirus and head and neck cancer. Am J Clin Oncol 32:535–539

    Article  PubMed  Google Scholar 

  16. Fliss MS, Usadel H, Caballero OL et al (2000) Facile detection of mitochondrial DNA mutations in tumors and bodily fluids. Science 287:2017–2019

    Article  PubMed  CAS  Google Scholar 

  17. Sidransky D (1997) Nucleic acid-based methods for the detection of cancer. Science 278:1054–1059

    Article  PubMed  CAS  Google Scholar 

  18. Rosas SL, Koch W, da Costa Carvalho MG et al (2001) Promoter hypermethylation patterns of p16, O6-methylguanine- DNA-methyltransferase, and death-associated protein kinase in tumors and saliva of head and neck cancer patients. Cancer Res 61:939–942

    PubMed  CAS  Google Scholar 

  19. Righini CA, de Fraipont F, Timsit JF et al (2007) Tumorspecific methylation in saliva: a promising biomarker for early detection of head and neck cancer recurrence. Clin Cancer Res 13:1179–1185

    Article  PubMed  CAS  Google Scholar 

  20. Franzmann EJ, Reategui EP, Pedroso F et al (2007) Soluble CD44 is a potential marker for the early detection of head and neck cancer. Cancer Epidemiol Biomarkers Prev 16:1348–1355

    Article  PubMed  CAS  Google Scholar 

  21. Nagler R, Bahar G, Shpitzer T et al (2006) Concomit antanalysis of salivary tumormarkers–a new diagnostic tool for oral cancer. Clin Cancer Res 12:3979–3984

    Article  PubMed  CAS  Google Scholar 

  22. Tavassoli M, Brunel N, Maher R et al (1998) p53 antibodies in the saliva of patients with squamous cell carcinoma of the oral cavity. Int J Cancer 78:390–391

    Article  PubMed  CAS  Google Scholar 

  23. Xie H, Onsongo G, Popko J et al (2008) Proteomics analysis of cells inwhole saliva from oral cancer patients via value-added three-dimensional peptide fractionation and tandem mass spectrometry. Mol Cell Proteomics 7:486–498

    PubMed  CAS  Google Scholar 

  24. Karpova MA, Moshkovskii SA, Toropygin IY, Archakov AI (2010) Cancer specific MALDI-TOF profiles of blood serum and plasma: biological meaning and perspectives. J Proteomics 73:537–551

    Article  PubMed  CAS  Google Scholar 

  25. Lee KD, Lee HS, Jeon CH (2011) Body fluid biomarkers for early detection of head and neck squamos cell carcinomas. Anticancer Res 31:1161–116

    PubMed  Google Scholar 

  26. Baker H, Patel V, Molinolo AA, Shillitoe EJ, Ensley JF, Yoo GH et al (2005) Proteome-wide analysis of head and neck squamous cell carinomas using laser-capture microdissecton and tandem mass spectrometry. Oral Oncol 41:183–199

    Article  PubMed  CAS  Google Scholar 

  27. El-Naggar AK, Mao L, Staekel G et al (2001) Genetic heterogeneity in saliva from patients with oral squamous carcinomas: implications in molecular diagnosis and screening. J Mol Diagn 3:164–170

    Article  PubMed  CAS  Google Scholar 

  28. Hu S, Arellano M, Boontheung P, Wang J, Zhou H, Jiang J, Elashoff D, Wei R, Loo JA, Wong DT (2008) Salivary proteomics for oral cancer biomarker discovery. Clin Cancer Res 14:6246–6252

    Article  PubMed  CAS  Google Scholar 

  29. Aizenbud D, Peri-Front Y, Nagler RM (2008) Salivary analysis and antioxidants in cleft lip and palate children. Arch Oral Biol 53:517–522

    Article  PubMed  CAS  Google Scholar 

  30. Navazesh M (1993) Methods for collecting saliva. Ann N Y Acad Sci 694:72–77

    Article  PubMed  CAS  Google Scholar 

  31. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  32. Gaal V, Mark L, Kiss P, Kustos I, Tamas A, Kocsis B, Lubics A, Nemeth V, Nemeth A, Lujber L, Pytel J, Toth G, Reglodi D (2008) Investigation of the effects of PACAP on the composition of tear and endolymph proteins. J Mol Neurosci 36:321–329

    Article  PubMed  CAS  Google Scholar 

  33. Baak JP, Path FR, Hermsen MA, Meijer G, Schmidt J, Janssen EA (2003) Genomics and proteomics in cancer. Eur J Cancer 39:1199–1215

    Article  PubMed  CAS  Google Scholar 

  34. Dowling P, Wormald R, Meleady P, Henry M, Curran A, Clynes M (2008) Analysis of the saliva proteome from patients with head and nack squamos cell carcinoma reveals differenes in abundance levels of proteins associated with tumour progression and metastasis. J Proteomics 71:168–175

    Article  PubMed  CAS  Google Scholar 

  35. Mahfouz ME, Rodrigo JP, Takes RP, Elsheikh MN, Rinaldo A, Brakenhoff RH, Ferlito A (2010) Current potential and limitations of molecular diagnostic methods in head and neck cancer. Eur Arch Otorhinolaryngol 267:851–860

    Article  PubMed  Google Scholar 

  36. Mydlarz WK, Hennessey PT, Califano JA (2010) Advances and perspectives in the molecular diagnosis of head and neck cancer. Expert Opin Med Diag 4:53–65

    Article  CAS  Google Scholar 

  37. Xia SH, Hu LP, Hu H, Ying WT, Xu X, Cai Y, Han YL, Chen BS, Wei F, Qian XH, Cai YY, Shen Y, Wu M, Wang MR (2002) Three isoforms of annexin I are preferentially expressed in normal esophageal epithelia but down-regulated in esophageal squamous cell carcinomas. Oncogene 21:6641–6648

    Article  PubMed  CAS  Google Scholar 

  38. Schaaij-Visser TB, Bremmer JF, Braakhuis BJ, Heck AJ, Slijper M, van der Waal I, Brakenhoff RH (2010) Evaluation of cornulin, keratin 4, keratin 13 expression and grade of dysplasia for predicting malignant progression of oral leukoplakia. Oral Oncol 46:123–127

    Article  PubMed  CAS  Google Scholar 

  39. Matsumoto I, Yamamoto M (2005) Differential expression of the keratin-4, -13,-14, -17 and transglutaminase 3 genes during the development of oral squamous cell carcinoma from leukoplakia. Oral Oncol 41:607–613

    Article  PubMed  Google Scholar 

  40. Pedrero JMG, Fernandez MP, Morgan RO, Zapatero AH, Gonzalez MV, Nieto CS, Rodrigo JP (2004) Annexin A1 down-regulation in head and neck cancer is associated with epithelial differentiation status. Am J Pathol 164:73–79

    Article  CAS  Google Scholar 

  41. Raynal P, Pollard HB (1994) Annexins: the problem of assessing the biological role for a gene family of multifunctional calcium-and-phosospholipid-binding proteins. Biochim Biophys Acta 1197:63–64

    PubMed  CAS  Google Scholar 

  42. Ohkura S, Kondoh N, Hada A, Arai M, Yamazaki Y, Sindoh M, Takahashi M, Mussunoor S, Murray GI (2008) The role of annexins in tumour development and progression. J Pathol 216:131–140

    Article  Google Scholar 

  43. Lin CY, Jeng YM, Chou HY, Hsu HC, Yuan RH, Chiang CP et al (2008) Nuclear localization of annexin A1 is a prognostic factor in oral squamous cell carcinoma. J Surg Oncol 97:544–550

    Article  PubMed  CAS  Google Scholar 

  44. Lo WY, Tsai MH, Tsai Y, Hua CH, Tsai FJ, Huang SY, Tsai CH, Lai CC (2007) Identification of overexpressed proteins in oral squamous cell carcinoma (OSCC) patients by clinical proteomic analysis. Clin Chim Acta 376:101–107

    Article  PubMed  CAS  Google Scholar 

  45. Makridakis M, Vlahou A (2010) Secretome proteomics for discovery of cancer biomarkers. J Proteomics 73:2291–2305

    Article  PubMed  CAS  Google Scholar 

  46. Schaaij-Visser T, Brakenhoff RH, Leemans CR, Heck AJR, Slijper M (2010) Protein biomarker discovery for head and neck cancer. J Proteomics 73:1790–1803

    Article  PubMed  CAS  Google Scholar 

  47. Schaaij-Visser TB, Graveland AP, Gauci S, Braakhuis BJ, Buijze M, Heck AJ et al (2009) Differential proteomics identifies protein biomarkers that predict local relapse of head and neck squamous cell carcinomas. Clin Cancer Res 15:7666–7675

    Article  PubMed  CAS  Google Scholar 

  48. Wadsworth JD, Somers K, Stack BC, Cazare L, Malik G, Adam BL, Wright GL, Semmes JO (2004) Identification of patients with head and neck cancer using serum protein profiles. Arch Otolaryngol Head Neck Surg 130:98–104

    Article  PubMed  Google Scholar 

  49. Wu W, Tang X, Hu W, Lotan R, Hong WK, Mao L (2002) Identification and validation of metastasis-associated proteins in head and neck cancer cell lines by two-dimensional electrophoresis and mass spectrometry. Clin Exp Metastasis 19:319–326

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The present work was supported by the Hungarian National Scientific Research Foundation (OTKA No. K72592, CNK78480), TIOP 1.3.1-10/1-2010-0008, TIOP 1.3.1-07/1, DDEK Kft., SciEx Kft., GVOP 0179 and PTE AOK KA 34039-11/2009 and KA 2011.

Author information

Authors and Affiliations

  1. Department of Oto-rhino-laryngology, Head and Neck Surgery, Medical School, University of Pecs, Pecs, Hungary

    Tamas Jarai, Andras Burian & Imre Gerlinger

  2. Institute of Biochemistry and Medical Chemistry, Medical School, University of Pecs, Szigeti str. 12, 7624, Pecs, Hungary

    Gabor Maasz, Agnes Bona, Eva Jambor & Laszlo Mark

Authors
  1. Tamas Jarai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gabor Maasz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andras Burian
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Agnes Bona
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Eva Jambor
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Imre Gerlinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Laszlo Mark
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Laszlo Mark.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jarai, T., Maasz, G., Burian, A. et al. Mass Spectrometry-Based Salivary Proteomics for the Discovery of Head and Neck Squamous Cell Carcinoma. Pathol. Oncol. Res. 18, 623–628 (2012). https://doi.org/10.1007/s12253-011-9486-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12253-011-9486-4

Keywords

Search

Navigation