Yu-Jen Jou1, Chun-Hung Hua2, Chia-Der Lin2, Chih-Ho Lai3, Su-Hua Huang4, Ming-Hsui Tsai2, Jung-Yie Kao5, Cheng-Wen Lin6. 1. Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung, Taiwan; Department of Biochemistry, College of Life Sciences, National Chung Hsing University, Taichung, Taiwan. 2. Department of Otolaryngology, China Medical University Hospital, Taichung, Taiwan. 3. Department of Microbiology, School of Medicine, China Medical University, Taichung, Taiwan. 4. Department of Biotechnology, College of Health Science, Asia University, Wufeng, Taichung, Taiwan. 5. Department of Biochemistry, College of Life Sciences, National Chung Hsing University, Taichung, Taiwan. 6. Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung, Taiwan; Department of Biotechnology, College of Health Science, Asia University, Wufeng, Taichung, Taiwan. Electronic address: cwlin@mail.cmu.edu.tw.
Abstract
BACKGROUND: Oral squamous cell carcinoma (OSCC) shows low 5-year survival; early treatment greatly reduces mortality and morbidity. Saliva is a non-invasive sample, with good potential to discover biomarkers for early detection. METHODS: NanoLC-MS/MS served to analyze saliva proteome from control subjects (n=35) and OSCC patients T1 (n=29), T2 (n=36), T3 (n=14) and T4 (n=21) stages. Identified biomarkers were verified by Western blot and ELISA assays. RESULTS: NanoLC-MS/MS analysis of salivary proteins between 10 and 15kDa identified S100A8, hemoglobin delta and gamma-G globin in T3 and T4 stage OSCC as well as S100A7 in T1 and T2 stage OSCC. Western blot and ELISA indicated positive correlation between salivary S100A8 increment and tumor size stage. High level of S100A8 appeared in 3.4, 13.9, 92.9, and 100% of saliva OSCC patients with T1, T2, T3, and T4 stages, respectively. Significant increase of salivary S100A7 was observed in 20.7% and 11.1% of those with T1 and T2, respectively. AUROC curve indicated high sensitivity, specificity and accuracy of S100A8-based ELISA as a detector. CONCLUSIONS: NanoLC-MS/MS, Western blot and ELISA manifested salivary S100A8 as a specific and sensitive marker for detection of OSCC patients. Salivary S100A8 protein could be applicable in developing OSCC diagnostics.
BACKGROUND:Oral squamous cell carcinoma (OSCC) shows low 5-year survival; early treatment greatly reduces mortality and morbidity. Saliva is a non-invasive sample, with good potential to discover biomarkers for early detection. METHODS: NanoLC-MS/MS served to analyze saliva proteome from control subjects (n=35) and OSCC patients T1 (n=29), T2 (n=36), T3 (n=14) and T4 (n=21) stages. Identified biomarkers were verified by Western blot and ELISA assays. RESULTS: NanoLC-MS/MS analysis of salivary proteins between 10 and 15kDa identified S100A8, hemoglobin delta and gamma-G globin in T3 and T4 stage OSCC as well as S100A7 in T1 and T2 stage OSCC. Western blot and ELISA indicated positive correlation between salivary S100A8 increment and tumor size stage. High level of S100A8 appeared in 3.4, 13.9, 92.9, and 100% of saliva OSCC patients with T1, T2, T3, and T4 stages, respectively. Significant increase of salivary S100A7 was observed in 20.7% and 11.1% of those with T1 and T2, respectively. AUROC curve indicated high sensitivity, specificity and accuracy of S100A8-based ELISA as a detector. CONCLUSIONS: NanoLC-MS/MS, Western blot and ELISA manifested salivary S100A8 as a specific and sensitive marker for detection of OSCC patients. Salivary S100A8 protein could be applicable in developing OSCC diagnostics.
Authors: M Castagnola; E Scarano; G C Passali; I Messana; T Cabras; F Iavarone; G Di Cintio; A Fiorita; E De Corso; G Paludetti Journal: Acta Otorhinolaryngol Ital Date: 2017-04 Impact factor: 2.124