Saliva-Based Multi-Omics Approaches for the Early Diagnosis of Oral Squamous Cell Carcinoma: A Comprehensive Review of Current Evidence and Clinical Perspectives
DOI:
https://doi.org/10.58600/eurjther3199Keywords:
early diagnosis, exosomes, liquid biopsy, oral squamous cell carcinoma, point-of-care systems, proteomics, salivary biomarkersAbstract
Objective: The aim of this review is to demonstrate that Oral Squamous Cell Carcinoma (OSCC) can be diagnosed early using saliva-based liquid biopsy focused on multi-omic biomarkers, and to evaluate the prognostic potential of this method as a non-invasive alternative to traditional tissue biopsies in long-term (longitudinal) follow-up.
Methods: A comprehensive literature review was conducted to synthesize current evidence regarding the diagnostic performance of multi-omic biomarker panels that include the biological composition of saliva, salivary proteomics, exosomal miRNAs, epigenetic profiles (DNA methylation), and oral microbiome dysbiosis. Additionally, the clinical applicability of integrating these multi-omic biomarkers with nanodiagnostic and Point-of-Care (POC) technologies was analyzed in conjunction with a comprehensive review of current translational challenges and critical approaches.
Results: Data from recent studies on the early detection of OSCC have shown that, when multiple biomarker panels are used, they achieve higher Area Under the Curve (AUC) values compared to individual biomarkers. In particular, exosomal miRNAs, epigenetic changes, and multi-omic data from saliva enable us to perform reliable risk assessments regarding disease prognosis and provide a clearer understanding of tumor mechanisms such as the epithelial-mesenchymal transition (EMT).
Conclusion: Saliva-based liquid biopsy approaches stand out as a promising, non-invasive option for the early diagnosis of oral cancers, monitoring prognosis, and evaluating treatment response. The integration of ctDNA, exosomal miRNA signatures, proteomic panels, and multi-omics approaches with artificial intelligence and machine learning algorithms can enhance diagnostic accuracy and support more personalized clinical decision-making processes. In the future, these technologies are expected to play a significant role in rapid, real-time, and cost-effective cancer screening in high-risk populations through artificial intelligence-supported systems integrated with point-of-care diagnostic methods.
References
[1] Towle R, Dickman CTD, MacLellan SA, Chen J, Prisman E, Guillaud M, Garnis C (2023) Identification of a serum-based microRNA signature that detects recurrent oral squamous cell carcinoma before it is clinically evident. Br J Cancer. 129(11):1810–1817. https://doi.org/10.1038/s41416-023-02405-9
[2] Bel’skaya LV, Sarf EA, Solomatin DV, Kosenok VK (2020) Diagnostic and prognostic value of salivary biochemical markers in oral squamous cell carcinoma. Diagnostics. 10(10):818. https://doi.org/10.3390/diagnostics10100818
[3] Bozyk N, Tang KD, Zhang X, Batstone M, Kenny L, Vasani S, Punyadeera C (2023) Salivary exosomes as biomarkers for early diagnosis of oral squamous cell carcinoma. Oral Oncol Rep. 6:100017. https://doi.org/10.1016/j.oor.2023.100017
[4] Shan J, Sun Z, Yang J, Xu J, Shi W, Wu Y, Fan Y, Li H (2019) Discovery and preclinical validation of proteomic biomarkers in saliva for early detection of oral squamous cell carcinomas. Oral Dis. 25(1):97–107. https://doi.org/10.1111/odi.12971
[5] Hsiao YC, Lin SY, Chien KY, Chen SF, Wu CC, Chang YT, Chi LM, Chu LJ, Chiang WF, Chien CY, Chang KP, Chang YS, Yu JS (2020) An immuno-MALDI mass spectrometry assay for the oral cancer biomarker, matrix metalloproteinase-1, in dried saliva spot samples. Anal Chim Acta. 1100:118–130. https://doi.org/10.1016/j.aca.2019.12.006
[6] Chang YT, Chu LJ, Liu YC, Chen CJ, Wu SF, Chen CH, Wang CY, Dash S, Chiang WF, Chiu KC, Gou HF, Chien CY, Yu JS (2020) Verification of saliva matrix metalloproteinase-1 as a strong diagnostic marker of oral cavity cancer. Cancers (Basel). 12(8):2273. https://doi.org/10.3390/cancers12082273
[7] Mehterov N, Vladimirov B, Sacconi A, Pulito C, Rucinski M, Blandino G, Sarafian V (2021) Salivary miR-30c-5p as Potential Biomarker for Detection of Oral Squamous Cell Carcinoma. Biomedicines. 9(9):1079. https://doi.org/10.3390/biomedicines9091079
[8] Romani C, Salviato E, Paderno A, Zanotti L, Ravaggi A, Deganello A, Berretti G, Gualtieri T, Marchini S, D'Incalci M, Mattavelli D, Piazza C, Bossi P, Romualdi C, Nicolai P, Bignotti E (2021) Genome-wide study of salivary miRNAs identifies miR-423-5p as promising diagnostic and prognostic biomarker in oral squamous cell carcinoma. Theranostics. 11(6):2987–2999. https://doi.org/10.7150/THNO.45157
[9] Gulati P, Singh AK, Yadav AK, Pasbola K, Pandey P, Sharma R, Thakar A, Solanki PR (2024) Nano-modified screen-printed electrode-based electrochemical immunosensors for oral cancer biomarker detection in undiluted human serum and saliva samples. Nanoscale Advances.6(2):705–721. https://doi.org/10.1039/D3NA00682D
[10] He L, Ping F, Fan Z, Zhang C, Deng M, Cheng B, Xia W (2020) Salivary exosomal miR-24-3p serves as a potential detective biomarker for oral squamous cell carcinoma screening. Biomed Pharmacother. 121:109553. https://doi.org/10.1016/j.biopha.2019.109553
[11] Heawchaiyaphum C, Pientong C, Phusingha P, Vatanasapt P, Promthet S, Daduang J, Teeramatwanich W, Kongyingyoes B, Chuerduangphui J, Ekalaksananan T (2018) Peroxiredoxin-2 and zinc-alpha-2-glycoprotein as potentially combined novel salivary biomarkers for early detection of oral squamous cell carcinoma using proteomic approaches. J Proteomics. 173:52–61. https://doi.org/10.1016/j.jprot.2017.11.022
[12] Lee LT, Wong YK, Hsiao HY, Wang YW, Chan MY, Chang KW (2018) Evaluation of saliva and plasma cytokine biomarkers in patients with oral squamous cell carcinoma. Int J Oral Maxillofac Surg. 47(6):699–707. https://doi.org/10.1016/j.ijom.2017.09.016
[13] Arantes LMRB, De Carvalho AC, Melendez ME, Centrone CC, Góis-Filho JF, Toporcov TN, Caly DN, Tajara EH, Goloni-Bertollo EM (2015) Validation of methylation markers for diagnosis of oral cavity cancer. Eur J Cancer. 51(5):632–641. https://doi.org/10.1016/j.ejca.2015.01.060
[14] Michailidou E, Tzimagiorgis G, Chatzopoulou F, Vahtsevanos K, Antoniadis K, Kouidou S, Markopoulos C, Antoniades K (2016) Salivary mRNA markers having the potential to detect oral squamous cell carcinoma segregated from oral leukoplakia with dysplasia. Cancer Epidemiol. 43:112–118. https://doi.org/10.1016/j.canep.2016.04.011
[15] Rodríguez-Zorrilla S, Lorenzo-Pouso AI, Fais S, Logozzi M, Mizzoni D, Di Raimo R, Giuliani M, García-García A, Pérez-Jardón A, Ortega KL, Martínez-González JM, Pérez-Sayáns M (2023) Increased plasmatic levels of exosomes are significantly related to relapse rate in patients with oral squamous cell carcinoma: a cohort study. Cancers (Basel). 15(23):5693. https://doi.org/10.3390/cancers15235693
[16] Bano A, Vats R, Verma D, Yadav P, Kamboj M, Bhardwaj R (2023) Exploring salivary exosomes as early predictors of oral cancer in susceptible tobacco consumers: noninvasive diagnostic and prognostic applications. J Cancer Res Clin Oncol. 149(17):15781–15793. https://doi.org/10.1007/s00432-023-05343-4
[17] Metgud R, Bajaj S (2016) Altered serum and salivary C-reactive protein levels in patients with oral premalignant lesions and oral squamous cell carcinoma. Biotech Histochem. 91(2):96–101. https://doi.org/10.3109/10520295.2015.1077393
[18] Dikova V, Jantus-Lewintre E, Bagan J (2021) Potential non-invasive biomarkers for early diagnosis of oral squamous cell carcinoma. J Clin Med. 10(8):1658. https://doi.org/10.3390/jcm10081658
[19] Song X, Yang X, Narayanan R, Shankar V, Ethiraj S, Wang X, Duan N, Ni YH, Hu Q, Zare RN (2020) Oral squamous cell carcinoma diagnosed from saliva metabolic profiling. Proc Natl Acad Sci U S A. 2020;117(28):16167–16173. https://doi.org/10.1073/pnas.2001395117/-/DCSupplemental
[20] Gupta A, Gupta S, Mahdi AA (2015) 1H NMR-derived serum metabolomics of leukoplakia and squamous cell carcinoma. Clin Chim Acta. 441:47–55. https://doi.org/10.1016/j.cca.2014.12.003
[21] Świątkowski W, Bakiera A, Rahnama-Hezavah M, Grywalska E, Niedzielski A, Korona-Głowniak I (2025) The oral microbiota change in oral cancer – a preliminary study. Ann Agric Environ Med. 32(3):353–360. https://doi.org/10.26444/aaem/209080
[22] van Dijk MC, Petersen JF, Raber-Durlacher JE, Epstein JB, Laheij AMGA (2024) Diversity and compositional differences in the oral microbiome of oral squamous cell carcinoma patients and healthy controls: a scoping review. Front Oral Health. 5:1366153. https://doi.org/10.3389/froh.2024.1366153
[23] Lim Y, Fukuma N, Totsika M, Kenny L, Morrison M, Punyadeera C (2018) The performance of an oral microbiome biomarker panel in predicting oral cavity and oropharyngeal cancers. Front Cell Infect Microbiol 8:267. https://doi.org/10.3389/fcimb.2018.00267
[24] Han Z, Hu Y, Lin X, Cheng H, Dong B, Liu X, Wu Y, Xu Y (2025) Systematic analyses uncover robust salivary microbial signatures and host-microbiome perturbations in oral squamous cell carcinoma. mSystems. 10(2):e01247-24. https://doi.org/10.1128/msystems.01247-24
[25] Chen YT, Chen HW, Wu CF, Chu LJ, Chiang WF, Wu CC, Yu YL, Tsai MH, Liang KH, Chang KP, Ou Yang MH (2017) Development of a multiplexed liquid chromatography multiple-reaction-monitoring mass spectrometry (LC-MRM/MS) method for evaluation of salivary proteins as oral cancer biomarkers. Mol Cell Proteomics. 16(5):799–811. https://doi.org/10.1074/mcp.M116.064758
[26] Yu JS, Chen YT, Chiang WF, Hsiao YC, Chu LJ, Seei LC, Wu CS, Tu HF, Liao YC, Chang KP, Wu CC, Lin CW, Liu CY, Chiou SH, Chang YT, Chien CY, Chang YL, Chang YC, Young DC, Pao CC, Chang JY, Hartwell L (2016) Saliva protein biomarkers to detect oral squamous cell carcinoma in a high-risk population in Taiwan. Proc Natl Acad Sci U S A. 113(41):11549–11554. https://doi.org/10.1073/pnas.1612368113
[27] Chu HW, Chang KP, Hsu CW, Chang IYF, Liu HP, Chen YT, Wu CC (2019) Identification of salivary biomarkers for oral cancer detection with untargeted and targeted quantitative proteomics approaches. Mol Cell Proteomics. 18(9):1796–1806. https://doi.org/10.1074/mcp.RA119.001530
[28] Patel A, Patel S, Patel P, Mandlik D, Patel K, Tanavde V (2022) Salivary exosomal miRNA-1307-5p predicts disease aggressiveness and poor prognosis in oral squamous cell carcinoma patients. Int J Mol Sci. 23(18):10639. https://doi.org/10.3390/ijms231810639
[29] Patel A, Patel P, Mandlik D, Patel K, Malaviya P, Johar K, Swamy KBS, Patel S, Tanavde V (2023) A novel 3-miRNA network regulates tumour progression in oral squamous cell carcinoma. Biomark Res. 11(1):64. https://doi.org/10.1186/s40364-023-00505-5
[30] Alafaria HAA, Jalal AS (2024) Novel DNA methylation biomarkers for early diagnosis of oral tongue squamous cell carcinoma (OTSCC). J Appl Genet. 65(3):541–548. https://doi.org/10.1007/s13353-024-00830-x
[31] Wu CC, Chu HW, Hsu CW, Chang KP, Liu HP (2015) Saliva proteome profiling reveals potential salivary biomarkers for detection of oral cavity squamous cell carcinoma. Proteomics. 15(19):3394–3404. https://doi.org/10.1002/pmic.201500157
[32] Blatt S, Kämmerer PW, Krüger M, Surabattula R, Thiem DGE, Dillon ST, Al-Nawas B, Libermann TA, Schuppan D (2023) High-multiplex aptamer-based serum proteomics to identify candidate serum biomarkers of oral squamous cell carcinoma. Cancers (Basel). 15(7):2071. https://doi.org/10.3390/cancers15072071
[33] Hazarika S, Mahesh S (2026) Fluid biopsy versus conventional biopsy – a revolutionary approach in oral cancer diagnosis: a systematic review and meta-analysis. Czas Stomatol. 79(1):73–82. https://doi.org/10.5114/jos.2026.160141
[34] Bitonti V, Gravili G, Berveglieri C, Familiari D, Cigni L, Franco R (2025) Salivary proteomics and its role in the early detection of oral cancer: a new frontier in noninvasive diagnostics. Open Med J. 13(1). https://doi.org/10.2174/0118742203418573251112070421
[35] Huang YY, You J-R, Lin PC (2025) Utility of quick salivary C-reactive protein assays in identifying early oral malignancy and supporting oral cancer detection. Int J Dent Res Allied Sci. 5(1):35–41. https://doi.org/10.51847/r3lrzgw8wn
[36] Mounts TE, Dillon JS, Slaughter IS (2024) Early Detection of Oral Squamous Cell Carcinoma Through Salivary Analysis. Undergraduate Research Scholars Capstone, Texas A&M University. https://hdl.handle.net/1969.1/203249
[37] Hanna K, Asiedu AL, Theurer T, Muirhead D, Speirs V, Oweis Y, Abu-Eid R (2024) Advances in Raman spectroscopy for characterising oral cancer and oral potentially malignant disorders. Expert Rev Mol Med. 26:e26. https://doi.org/10.1017/erm.2024.26
[38] Bhaskar S, Umrao S, Lee HK, Tibbs J, Bacon A, Shepherd S, Ayupova T, Ciloglu FU, Liu L, Tan A, Chen WC, Nguyen MTT, Scannell MG, Aygun U, Parlatan U, Zhang C, Kohli M, Adami GR, Badar W, Gaba RC, Mansfield A, Schwartz J, Xing W, Demirci U, Cunningham BT (2026) Point of care molecular cancer diagnostics. Lab Chip. 26(10):2948–3001. https://doi.org/10.1039/D5LC01014D
[39] Pierfelice TV, D’Amico E, Cinquini C, Iezzi G, D’Arcangelo C, D’Ercole S, Petrini M (2024) The diagnostic potential of non-invasive tools for oral cancer and precancer: a systematic review. Diagnostics (Basel). 14(18):2033. https://doi.org/10.3390/diagnostics14182033
[40] Ramani RS (2025) Machine Learning and Fluorescence In Vivo Confocal Microscopy for the Early Detection of Oral Potentially Malignant Disorders and Oral Cancer. Ph.D. thesis, The University of Melbourne. https://minerva-access.unimelb.edu.au/handle/11343/371230
[41] Warnakulasuriya S, Patil S (2025) Salivary and serum liquid biopsy biomarkers for HPV-associated oral and oropharyngeal cancer: a narrative review. J Clin Med. 14(21):7598. https://doi.org/10.3390/jcm14217598
[42] Rashid S, Puttagunta P, Pamulapati S, Yang J, Pocha S, Saba NF, Teng Y (2024) Leveraging saliva for insights into head and neck cancer. Int J Mol Sci. 25(24):13514. https://doi.org/10.3390/ijms252413514
[43] Zehra T, Koopaie M, Fatima N, Rashid G, Hasan I, Siddiqui Z (2026) Artificial intelligence-based miRNA analysis for precision oncology: diagnostic and prognostic insights. Front Mol Biosci. 13:1749586. https://doi.org/10.3389/fmolb.2026.1749586
[44] Srivathsa S, Vinjamuri S, Narayanan P, Reddy SDN (2026) Decoding microRNAs as potential biomarkers and therapeutic targets in head and neck cancer. Oncol Transl Med. (Epub ahead of print articles) https://doi.org/10.1097/ot9.0000000000000142
[45] Mali SB (2023) Molecular screening of head neck cancer. Oral Oncol. 144:106481. https://doi.org/10.1016/j.oraloncology.2023.106481
[46] Morelli I, Ghirardini C, Faccani L, Casanova C, Fernandez IJ, Tamberi S (2025) Liquid biopsy and circulating biomarkers in head and neck cancer: advancing non-invasive detection and tailored management. Cancers (Basel). 17(24):3974. https://doi.org/10.3390/cancers17243974
[47] Zhao LX, Wang ET, Jia WH, Wang MH, Pan Z, Dong L, Wen Y, Xu X, Xu X (2026) Emerging salivary exosome nanodiagnostics for early oral squamous cell carcinoma: platforms, biomarkers, and future perspectives. Int J Nanomedicine. 21:577306. https://doi.org/10.2147/IJN.S577306
[48] Hsu PC, Huang JH, Tsai CC, Lin YH, Kuo CY (2025) Early molecular diagnosis and comprehensive treatment of oral cancer. Curr Issues Mol Biol. 47(6):452. https://doi.org/10.3390/cimb47060452
[49] Ebrahim NAA, Soliman SMA, Farghaly TA (2025) Saliva-based molecular diagnostics in oral squamous cell carcinoma (OSCC): a non-invasive frontier in oncology. Eur Arch Otorhinolaryngol. https://doi.org/10.1007/s00405-025-09934-4
[50] Kumar R, Keshari P, Kant A, Kumar A, Gupta R, Yadav SC (2024) Insights into noninvasive approaches for the early detection of cancer with a special focus on nanodiagnostics. In: Nano Biosensors for Non-Invasive Diagnosis of Cancer. Ingram Publisher Services UK, Bristol, pp 4-1–4-42. https://doi.org/10.1088/978-0-7503-6234-4ch4
[51] Verma A, Arqam M, Fraiwan A (2025) Advances in impedimetric biosensors: current applications and future directions. Micromachines (Basel). 16(11):1244. https://doi.org/10.3390/mi16111244
[52] Sánchez-Tirado E, Agüí L, González-Cortés A, Campuzano S, Yáñez-Sedeño P, Pingarrón JM (2023) Electrochemical (bio)sensing devices for human-microbiome-related biomarkers. Sensors (Basel). 23(2):837. https://doi.org/10.3390/s23020837
[53] Prasad M, Sekar R, Priya MDL, Varma SR, Karobari MI (2024) A new perspective on diagnostic strategies concerning the potential of saliva-based miRNA signatures in oral cancer. Diagn Pathol. 19(1):147. https://doi.org/10.1186/s13000-024-01575-1
[54] Li Y, Ou Y, Fan K, Liu G (2024) Salivary diagnostics: opportunities and challenges. Theranostics. 14(18):6969–6990. https://doi.org/10.7150/thno.100600
[55] Görmez Ö, Aydemir ME, Çelebi E, Yıldırım D (2020) Oral skuamöz hücreli karsinom olgularının beş yıllık literatür incelemesi. Osmangazi Journal of Medicine. 42(5):229–232. https://doi.org/10.20515/otd.772066
Downloads
Published
How to Cite
Issue
Section
Categories
License
Copyright (c) 2026 Perya Pelin Özsöyler Bozan, Mustafa Örkmez
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
The content of this journal is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
