采用纳升液相色谱-串联质谱法研究 S100A8 作为口腔鳞状细胞癌潜在唾液生物标志物的研究。
S100A8 as potential salivary biomarker of oral squamous cell carcinoma using nanoLC-MS/MS.
机构信息
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.
Department of Otolaryngology, China Medical University Hospital, Taichung, Taiwan.
出版信息
Clin Chim Acta. 2014 Sep 25;436:121-9. doi: 10.1016/j.cca.2014.05.009. Epub 2014 May 24.
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.
背景
口腔鳞状细胞癌（OSCC）的五年生存率较低；早期治疗可大大降低死亡率和发病率。唾液是一种非侵入性样本，具有发现早期检测生物标志物的巨大潜力。
方法
采用纳升液相色谱-串联质谱（nanoLC-MS/MS）分析对照组（n=35）和 OSCC 患者 T1（n=29）、T2（n=36）、T3（n=14）和 T4（n=21）期的唾液蛋白质组。通过 Western blot 和 ELISA 检测验证鉴定出的生物标志物。
结果
在 10 至 15kDa 的唾液蛋白之间进行的 nanoLC-MS/MS 分析，鉴定出 T3 和 T4 期 OSCC 中的 S100A8、血红蛋白 delta 和 gamma-G 球蛋白，以及 T1 和 T2 期 OSCC 中的 S100A7。Western blot 和 ELISA 表明唾液 S100A8 增量与肿瘤大小分期之间存在正相关。在 T1、T2、T3 和 T4 期的 OSCC 患者中，分别有 3.4%、13.9%、92.9%和 100%的患者唾液中 S100A8 水平较高。在 T1 和 T2 期的患者中，分别有 20.7%和 11.1%的患者唾液中 S100A7 显著增加。ROC 曲线表明基于 ELISA 的 S100A8 检测具有高灵敏度、特异性和准确性。
结论
nanoLC-MS/MS、Western blot 和 ELISA 表明唾液 S100A8 是检测 OSCC 患者的特异性和敏感性标志物。唾液 S100A8 蛋白可应用于开发 OSCC 诊断。
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