Use of Combination Proteomic Analysis to Demonstrate Molecular Similarity of Head and Neck Squamous Cell Carcinoma Arising From Different Subsites

Abstract

BACKGROUND Squamous cell carcinomas arising from various subsites within the head and neck (HNSCC), while histologically identical, have substantial differences in survival and recurrence rates. Controversy exists as to whether this reflects physical differences between subsites or fundamental molecular heterogeneity. In this study, we used two proteomic approaches to evaluate HNSCCs for differences in protein expression between oral cavity, oropharynx, larynx and hypopharynx subsites. METHODS A tissue microarray (TMA) was constructed consisting of 71 patients with HNSCC. This TMA was queried for expression of 4 cell- cycle and regulatory proteins chosen a priori for their known roles in cancer, using Automated Quantitative Analysis of protein expression (AQUA). Frozen tissue samples from 14 patients with histologically confirmed HNSCC were enriched for tumor and normal tissue by laser capture microdissection. Total protein was extracted, analyzed by 2D-difference gel electrophoresis (2D-DIGE) with saturation dye labeling, and evaluated for differential protein expression between subsites. RESULTS AQUA analysis revealed no difference between subsite for cyclin D1, p53, Rb, or p14 expression. The 2D-DIGE study was based on 28 gels (14 cancer, 14 adjacent normal) and 732 spots were identified as matching across >90% of gels. Statistical analysis detected no significant differences in protein expression between subsites. CONCLUSIONS Observed differences in outcomes between HNSCCs from different subsites may not reflect differences in tumor biology between subsites. Rather, it is possible that observed clinical heterogeneity among HNSCCs may be based on other factors, such as viral versus chemical carcinogenesis.

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