Potential role of nuclear magnetic resonance spectroscopy to identify salivary metabolite alterations in patients with head and neck cancer

Mikkonen,Jopi J.W.; Singh,Surya P.; Akhi,Ramin; Salo,Tuula; Lappalainen,Reijo; González‑Arriagada,Wilfredo A.; Ajudarte Lopes,Márcio; Kullaa,Arja M.; Myllymaa,Sami

doi:10.3892/ol.2018.9419

Potential role of nuclear magnetic resonance spectroscopy to identify salivary metabolite alterations in patients with head and neck cancer

Authors:
- Jopi J.W. Mikkonen
- Surya P. Singh
- Ramin Akhi
- Tuula Salo
- Reijo Lappalainen
- Wilfredo A. González‑Arriagada
- Márcio Ajudarte Lopes
- Arja M. Kullaa
- Sami Myllymaa
View Affiliations

Affiliations: SIB Labs, University of Eastern Finland, FI‑70211 Kuopio, Finland, Institute of Dentistry, University of Eastern Finland, FI‑70211 Kuopio, Finland, Research Unit of Oral Health Sciences, University of Oulu, FI‑90014 Oulu, Finland, Cancer and Translational Medicine Research Unit, University of Oulu, FI‑90014 Oulu, Finland, Oral Pathology and Diagnosis, School of Dentistry, Universidad de Valparaiso, Valparaiso, Región de Valparaíso 2360004, Chile, Department of Oral Diagnosis, School of Dentistry, State University of Campinas, Sao Paulo CEP 13414‑018, Brazil
Published online on: September 7, 2018 https://doi.org/10.3892/ol.2018.9419
Pages: 6795-6800

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The analysis of the salivary metabolomic profile may offer an early phase approach to assess the changes associated with a wide range of diseases including head and neck cancer. The aim of the present study was to investigate the potential of nuclear magnetic resonance (NMR) spectroscopy for detecting the salivary metabolic changes associated with head and neck squamous cell carcinoma (HNSCC). Unstimulated whole‑mouth saliva samples collected from HNSCC patients (primary tumour was located either in the larynx or in the oral cavity) and healthy controls were analysed by 1H‑NMR spectroscopy. Reliably identified salivary metabolites were quantified and the determined concentration values were compared group‑wise using a Mann‑Whitney U‑test. Multivariate discrimination function analysis (DFA) was conducted to identify such a combination of metabolites, when considered together, that gives maximum discrimination between the groups. HNSCC patients exhibited significantly increased concentrations of 1,2‑propanediol (P=0.032) and fucose (P=0.003), while proline levels were significantly decreased (P=0.043). In the DFA model, the most powerful discrimination was achieved when fucose, glycine, methanol and proline were considered as combined biomarkers, resulting in a correct classification rate of 92.1%, sensitivity of 87.5% and specificity of 93.3%. To conclude, NMR spectrometric analysis was revealed to be a feasible approach to study the metabolome of saliva that is sensitive to metabolic changes in HNSCC and straightforward to collect in a non‑invasive manner. Salivary fucose was of particular interest and therefore, controlled longitudinal studies are required to assess its clinical relevance as a diagnostic biomarker in HNSCC.

View Figures

View References

1	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2015. View Article : Google Scholar : PubMed/NCBI
2	Abbey LM, Kaugars GE, Gunsolley JC, Burns JC, Page DG, Svirsky JA, Eisenberg E, Krutchkoff DJ and Cushing M: Intraexaminer and interexaminer reliability in the diagnosis of oral epithelial dysplasia. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 80:188–191. 1995. View Article : Google Scholar : PubMed/NCBI
3	Liu J and Duan Y: Saliva: A potential media for disease diagnostics and monitoring. Oral Oncol. 48:569–577. 2012. View Article : Google Scholar : PubMed/NCBI
4	Zhang CZ, Cheng XQ, Li JY, Zhang P, Yi P, Xu X and Zhou: Saliva in the diagnosis of diseases. Int J Oral Sci. 8:133–137. 2016. View Article : Google Scholar : PubMed/NCBI
5	Yoshizawa JM, Schafer CA, Schafer JJ, Farrell JJ, Paster BJ and Wong DT: Salivary biomarkers: Toward future clinical and diagnostic utilities. Clin Microbiol Rev. 26:781–791. 2013. View Article : Google Scholar : PubMed/NCBI
6	Cuevas-Córdoba B and Santiago-García J: Saliva: A fluid of study for OMICS. OMICS. 18:87–97. 2014. View Article : Google Scholar : PubMed/NCBI
7	Guerra EN, Acevedo AC, Leite AF, Gozal D, Chardin H and De Luca Canto G: Diagnostic capability of salivary biomarkers in the assessment of head and neck cancer: A systematic review and meta-analysis. Oral Oncol. 51:805–818. 2015. View Article : Google Scholar : PubMed/NCBI
8	Patel S and Ahmed S: Emerging field of metabolomics: Big promise for cancer biomarker identification and drug discovery. J Pharm Biomed Anal. 107:63–74. 2015. View Article : Google Scholar : PubMed/NCBI
9	Mikkonen JJW, Herrala M, Soininen P, Lappalainen R, Tjäderhane L, Seitsalo H, Niemelä R, Salo T, Kullaa A and Myllymaa S: Metabolic profiling of saliva in patients with primary Sjögren's syndrome. Metabolomics. 3:1282013.
10	Spielmann N and Wong DT: Saliva: Diagnostics and therapeutic perspectives. Oral Dis. 17:345–354. 2011. View Article : Google Scholar : PubMed/NCBI
11	Bonne NJ and Wong DT: Salivary biomarker development using genomic, proteomic and metabolomic approaches. Genome Med. 4:822012. View Article : Google Scholar : PubMed/NCBI
12	Winck FV, Ribeiro Prado AC, Domingues Ramos R, Ling LY, Riaño-Pachón DM, Rivera C, Brandão TB, Gouvea AF, Santos-Silva AR, Coletta RD and Paes Leme AF: Insights into immune responses in oral cancer through proteomic analysis of saliva and salivary extracellular vesicles. Sci Rep. 5:163052015. View Article : Google Scholar : PubMed/NCBI
13	Zlotogorski-Hurvitz A, Dayan D, Chaushu G, Salo T and Vered M: Morphological and molecular features of oral fluid-derived exosomes: Oral cancer patients versus healthy individuals. J Cancer Res Clin Oncol. 142:101–110. 2016. View Article : Google Scholar : PubMed/NCBI
14	Günther H: NMR spectroscopy: Basic principles, concepts, and applications in chemistry. 2 edition. John Wiley and Sons Ltd.; Chichester: 1995
15	González-Arriagada WA, Ramos LM, Silva AA, Vargas PA, Coletta RD, Bingle L and Lopes MA: Salivary BPIFA1 (SPLUNC1) and BPIFA2 (SPLUNC2 A) are modified by head and neck cancer radiotherapy. Oral Surg Oral Med Oral Pathol Oral Radiol. 119:48–58. 2015. View Article : Google Scholar : PubMed/NCBI
16	Navazesh M: Methods for collecting saliva. Ann N Y Acad Sci. 694:72–77. 1993. View Article : Google Scholar : PubMed/NCBI
17	Silwood CJ, Lynch E, Claxson AW and Grootveld MC: 1H and (13)C NMR spectroscopic analysis of human saliva. J Dent Res. 81:422–427. 2002. View Article : Google Scholar : PubMed/NCBI
18	Dame ZT, Aziat F, Mandal RS, Krishnamurthy R, Bouatra S, Borzouie S, Guo AC, Sajed T, Deng L, Lin H, et al: The human saliva metabolome. Metabolomics. 11:1864–1883. 2015. View Article : Google Scholar
19	Soininen P, Haarala J, Vepsäläinen J, Niemitz M and Laatikainen R: Strategies for organic impurity quantification by 1H NMR spectroscopy: Constrained total-line-shape fitting. Anal Chim Acta. 542:178–185. 2005. View Article : Google Scholar
20	Ma B, Simala-Grant JL and Taylor DE: Fucosylation in prokaryotes and eukaryotes. Glycobiology. 16:158R–184R. 2006. View Article : Google Scholar : PubMed/NCBI
21	Miyoshi E, Moriwaki K and Nakagawa T: Biological function of fucosylation in cancer biology. J Biochem. 143:725–729. 2008. View Article : Google Scholar : PubMed/NCBI
22	Fernández-Rodríguez J, de la Cadena Páez M, Martínez-Zorzano VS and Rodríguez-Berrocal FJ: Fucose levels in sera and in tumours of colorectal adenocarcinoma patients. Cancer Lett. 121:147–153. 1997. View Article : Google Scholar : PubMed/NCBI
23	Shah M, Telang S, Raval G, Shah P and Patel PS: Serum fucosylation changes in oral cancer and oral precancerous conditions: Alpha-L-fucosidase as a marker. Cancer. 113:336–346. 2008. View Article : Google Scholar : PubMed/NCBI
24	Manjula S, Monteiro F, Aroor Rao A, Rao S, Annaswamy R and Rao A: Assessment of serum L-fucose in brain tumor cases. Ann Indian Acad Neurol. 13:33–36. 2010. View Article : Google Scholar : PubMed/NCBI
25	Listinsky JJ, Siegal GP and Listinsky CM: The emerging importance of α-L-fucose in human breast cancer: A review. Am J Transl Res. 3:292–322. 2011.PubMed/NCBI
26	Shetlar MR, Foster JV, Kelly KH, Shetlar CL, Bryan RS and Everett MR: The serum polysaccharide level in malignancy and in other pathological conditions. Cancer Res. 9:515–519. 1949.PubMed/NCBI
27	Sawke NG and Sawke GK: Serum fucose level in malignant diseases. Indian J Cancer. 47:452–457. 2010. View Article : Google Scholar : PubMed/NCBI
28	Shetty RK, Bhandary SK and Kali A: Significance of serum L-fucose glycoprotein as cancer biomarker in head and neck malignancies without distant metastasis. J Clin Diagn Res. 7:2818–2820. 2013.PubMed/NCBI
29	Parwani RN and Parwani SR: Quantitative evaluation of serum fucose in oral squamous cell carcinoma patients. J Can Res Ther. 7:143–147. 2011. View Article : Google Scholar
30	Rai NP, Anekar J, Shankara Shivaraja YM, Divakar DD, Al Kheraif AA, Ramakrishnaiah R, Sebastian R, Raj AC, Al-Hazmi A and Mustafa SM: Comparison of serum fucose levels in leukoplakia and oral cancer patients. Asian Pac J Cancer Prev. 16:7497–7500. 2015. View Article : Google Scholar : PubMed/NCBI
31	Chugh S, Gnanapragassam VS, Jain M, Rachagani S, Ponnusamy MP and Batra SK: Pathobiological implications of mucin glycans in cancer: Sweet poison and novel targets. Biochim Biophys Acta. 1856:211–225. 2015.PubMed/NCBI
32	Rajpura KB, Patel PS, Chawda JG and Shah RM: Clinical significance of total and lipid bound sialic acid levels in oral pre-cancerous conditions and oral cancer. J Oral Pathol Med. 34:263–267. 2005. View Article : Google Scholar : PubMed/NCBI
33	Sanjay PR, Hallikeri K and Shivashankara AR: Evaluation of salivary sialic acid, total protein, and total sugar in oral cancer: A preliminary report. Indian J Dent Res. 19:288–291. 2008. View Article : Google Scholar : PubMed/NCBI
34	Sawhney H and Kumar CA: Correlation of serum biomarkers (TSA & LSA) and epithelial dysplasia in early diagnosis of oral precancer and oral cancer. Cancer Biomark. 10:43–49. 2011. View Article : Google Scholar : PubMed/NCBI
35	Taqi SA: Clinical evaluation of total and lipid bound sialic acid levels in oral precancer and oral cancer. Indian J Med Paediatr Oncol. 33:36–41. 2012. View Article : Google Scholar : PubMed/NCBI
36	Dhakar N, Astekar M, Jain M, Saawarn S and Saawarn N: Total sialic acid, total protein and total sugar levels in serum and saliva of oral squamous cell carcinoma patients: A case control study. Dent Res J (Isfahan). 10:343–347. 2013.PubMed/NCBI
37	Dadhich M, Prabhu V, Pai VR, D'Souza J, Harish S and Jose M: Serum and salivary sialic acid as a biomarker in oral potentially malignant disorders and oral cancer. Indian J Cancer. 51:214–218. 2014. View Article : Google Scholar : PubMed/NCBI
38	Mustafa A, Gupta S, Hudes GR, Egleston BL, Uzzo RG and Kruger WD: Serum amino acid levels as a biomarker for renal cell carcinoma. J Urol. 186:1206–1212. 2011. View Article : Google Scholar : PubMed/NCBI
39	Tiziani S, Lopes V and Günther UL: Early stage diagnosis of oral cancer using 1H NMR-based metabolomics. Neoplasia. 11(269–276): 4p2009.
40	Liang S, Sanchez-Espiridion B, Xie H, Ma J, Wu X and Liang D: Determination of proline in human serum by a robust LC-MS/MS method: Application to identification of human metabolites as candidate biomarkers for esophageal cancer early detection and risk stratification. Biomed Chromatogr. 29:570–577. 2015. View Article : Google Scholar : PubMed/NCBI
41	Humphrey SP and Williamson RT: A review of saliva: Normal composition, flow, and function. J Prosthet Dent. 85:162–169. 2001. View Article : Google Scholar : PubMed/NCBI
42	Tiwari M: Science behind human saliva. J Nat Sci Biol Med. 2:53–58. 2011. View Article : Google Scholar : PubMed/NCBI
43	Koneru S and Tanikonda R: Salivaomics-A promising future in early diagnosis of dental diseases. Dent Res J (Isfahan). 11:11–15. 2014.PubMed/NCBI
44	Hu S, Gao K, Pollard R, Arellano-Garcia M, Zhou H, Zhang L, Elashoff D, Kallenberg CG, Vissink A and Wong DT: Preclinical validation of salivary biomarkers for primary Sjögren's syndrome. Arthritis Care Res (Hoboken). 62:1633–1638. 2010. View Article : Google Scholar : PubMed/NCBI
45	Mikkonen JJ, Singh SP, Herrala M, Lappalainen R, Myllymaa S and Kullaa AM: Salivary metabolomics in the diagnosis of oral cancer and periodontal diseases. J Periodontal Res. 51:431–437. 2016. View Article : Google Scholar : PubMed/NCBI
46	Bigler LR, Streckfus CF, Copeland L, Burns R, Dai X, Kuhn M, Martin P and Bigler SA: The potential use of saliva to detect recurrence of disease in women with breast carcinoma. J Oral Pathol Med. 31:421–431. 2002. View Article : Google Scholar : PubMed/NCBI
47	Zhang L, Xiao H, Zhou H, Santiago S, Lee JM, Garon EB, Yang J, Brinkmann O, Yan X, Akin D, et al: Development of transcriptomic biomarker signature in human saliva to detect lung cancer. Cell Mol Life Sci. 69:3341–3350. 2012. View Article : Google Scholar : PubMed/NCBI
48	Xiao H, Zhang L, Zhou H, Lee JM, Garon EB and Wong DT: Proteomic analysis of human saliva from lung cancer patients using two-dimensional difference gel electrophoresis and mass spectrometry. Mol Cell Proteomics. 11(M111): 0121122012.PubMed/NCBI
49	Sugimoto M, Wong DT, Hirayama A, Soga T and Tomita M: Capillary electrophoresis mass spectrometry-based saliva metabolomics identified oral, breast and pancreatic cancer-specific profiles. Metabolomics. 6:78–95. 2010. View Article : Google Scholar : PubMed/NCBI
50	Lau C, Kim Y, Chia D, Spielmann N, Eibl G, Elashoff D, Wei F, Lin YL, Moro A, Grogan T, et al: Role of pancreatic cancer-derived exosomes in salivary biomarker development. J Biol Chem. 288:26888–26897. 2013. View Article : Google Scholar : PubMed/NCBI
51	Grant MM: What do'omic technologies have to offer periodontal clinical practice in the future? J Periodontal Res. 47:2–14. 2012. View Article : Google Scholar : PubMed/NCBI