Abstract

Serum proteomic analysis can be a valuable approach for the discovery of protein biomarkers for early detection or monitoring of a disease. In this study, two analytical methods were compared for quantification of serum proteins in patients with oral cancer. In the first approach, we quantified serum proteins between oral squamous cell carcinoma (OSCC) and healthy control subjects by performing in-solution digestion of serum proteins, isobaric tags for relative and absolute quantification (iTRAQ) labeling of the resulting peptides, strong cation exchange (SCX) fractionation of labeled peptides and finally capillary liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis of the peptides. In the second approach, we first separated serum proteins with SDS-PAGE. The gel-separated proteins were then digested with trypsin and the resulting peptides were labeled with iTRAQ and analyzed with LC-MS/MS for protein quantification. A total of 319 serum proteins were quantified with the first proteomic approach whereas a total of 281 proteins were quantified by the second proteomic approach. Most of the proteins were identified and quantified by both approaches, suggesting that these methods are similarly effective for serum proteome analysis. This study provides compelling evidence that quantitative serum proteomic analysis of OSCC is a valuable approach for identifying differentially expressed proteins in cancer patients’ circulation systems that may be used as potential biomarkers for disease detection. Further validation in large oral cancer patient populations may lead to a simple and low invasive clinical tool for OSCC diagnosis or monitoring.

Highlights

  • Oral cancer is a subgroup of head and neck cancer that affects many regions of the oral cavity, such as the lips, tongue, salivary glands, and gums

  • Using similar quantitative proteomics approaches, we have previously identified serum protein biomarkers for classification of oral cancer patients with lymph node metastasis [13] and revealed that the cAMP response element-binding protein 1 (CREB1) pathway is activated in oral cancer stem-like cells [14]

  • Two analytical methods were compared in this study for the discovery of putative serum protein biomarkers of oral cancer

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Summary

Introduction

Oral cancer is a subgroup of head and neck cancer that affects many regions of the oral cavity, such as the lips, tongue, salivary glands, and gums. It is considered as the sixth most common cancer, accounting for nearly 3% of the total cancer burden and results in 128,000 annual deaths globally [1,2]. The most common type of oral cancer is oral squamous cell carcinoma (OSCC), which makes up 90%. A number of studies were performed to investigate the clinical utility of tumor antigens, such as carcinoembryonic antigen (CEA), CA-50, CA19-9 and squamous cell carcinoma antigen (SCCA), for OSCC detection [6,7,8,9]

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