Abstract
Purpose The purpose of this study is to identify biomarkers from the salivary proteome for early detection/prognostication of Oral Squamous Cell Carcinoma (OSCC). Materials and methods Patients diagnosed with OSCC and premalignant lesions were recruited for the study and healthy volunteers were used as controls. Pre-treatment saliva samples were collected and processed according to standard protocols. Proteomic profiling of normal and diseased saliva was carried out by multiple methods; mass spectrometric analysis following iTRAQ labeling methods was used for differential profiling. The data from fractions were analyzed using the commercially available MASCOT and SEQUEST software. Results Processing of the salivary supernatant was standardized for the purpose of depleting abundant proteins; starch, trisacryl resin and MARS column based depletion of amylase and albumin respectively was carried out. MS analysis on normal saliva and subsequent comparison with existing markers in the database identified 213 novel proteins. The proteomic profile of the tumor group (buccal mucosal cancer) with metastatic lymph nodes was compared to matched normal controls (N = 5 each). A total of 150 differential proteins were obtained after analysis by the MASCOT software. These proteins included Profilin, Cofilin, S100A9 and MMP9 identified previously in the saliva of head and neck cancer patients along with others associated with epithelial mesenchymal transition and metastasis such as Vimentin. Ingenuity Pathway Analysis (IPA) identified multiple networks which included proteins involved in cellular movement, cell–cell signaling, cell growth and proliferation. Further studies to validate these markers in tissues/saliva are currently ongoing. Proteomic profiling of the other study groups are also underway to identify biomarkers that specify the clinical status of the patients. Discussion Proteomic profiling of the salivary biomarkers contributed towards identifying novel markers in the normal subjects and HNSCC patients with lymphatic metastasis; validation in the saliva/tissues of the patients will enable assessment of their clinical application.
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