Despite considerable development in the therapeutic repertoire for managing cancer-related malignancies, head and neck cancer mortality has not significantly improved. The burden of HNSCC fluctuates across countries and has been associated with exposure to tobacco-derived carcinogens, excessive alcohol consumption or combinations. Due to late detection, patients often present with oral pre-malignant lesions which have progressed to an advanced stage of HNSCC. In this study, the samples were from a male cohort as generally, men are at two to four-fold higher risk than women with over 90% of HNSCCs arising in the upper aerodigestive tract. Therefore, the purpose of this thesis was to identify HNSCC biomarkers in males associated within defined anatomical region (tongue) and causative agents, specific to smoking. An iTRAQ proteomic approach was used to profile protein changes in matched normal and tumour samples from male non-smoking (n=6) and smoking patients (n=6) with tongue carcinomas revealing identification of potential targets specific to cancer. Samples were subjected to liquid nitrogen cryo-pulverisation and protein determination. Protein extracts from the same category were pooled, trypsin digested and iTRAQ 4-plex labelled. Data was generated by 2D-LC/MS on an Orbitrap Fusion and significantly changed proteins (median ± SD) were subject to bioinformatics appraisal. A total of 3426 proteins were identified and quantified by proteomic analysis. Comparison of non-smoker tumour (NS:T) with smoker tumour (S:T) distinguished 64 proteins that were upregulated and 62 downregulated, S:T vs S:N categorised 349 proteins up- and 395 down-regulated respectively and NS:T vs NS:N identified 469 proteins up- and 431 down-regulated, respectively. Arginase-1 (ARG1), Keratin Type-2 Cytoskeletal 8 (KRT8), Lipocalin-1 (LCN1) and DNA replication licensing factor MCM2 (MCM2) were identified as biologically associated with smoking compared to non-smoking, providing viable targets for verification by immunochemical methods which further supported the proteomic data. Overall, the project demonstrated the importance of using matched biopsies with good clinicopathological data for experimental design and provided a set of unique targets for a more expanded verification study.