A Label-Free Quantitation Method by MS/MS Spectral Counting for Urine Biomarker Discovery of Bladder Cancer

• Main Authors: Cheng Han Tsai, 蔡承翰
• Other Authors: Y. T. Chen
• Format: Others
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/64463089287960920966

碩士 === 長庚大學 === 生物醫學研究所 === 99 === Bladder cancer is the second most common malignant carcinoma in the urogenital disease. The newly diagnose and death cases increase every year. The current diagnostic methods are invasive and expensive in clinical. Development of an non-invasive method for detection of bladder cancer is required. Although many bladder cancer biomarkers have been reported, but none of them have shown sufficient sensitivity and specificity. Therefore, discovery of bladder cancer biomarker in urine is important. In this study, we used the high-abundant proteins depletion technology to remove the high-abundant proteins in urine that may cause the ion suppression effect in LC-MS/MS analysis. The depletion of high-abundant proteins was performed using the MARS Hu-14 column and Prospectrum librariesTM beads. The data indicated that depletion of high abundant urine protein enhanced the detection of low-abundant proteins and decreased the dynamic range of urine proteins. Furthermore, up-expressed urine proteins of bladder cancer and proteins only present in cancer were compared with secretome datasets of bladder cancer cell lines, HUPO HPPP plasma dataset and sys-body fluid urine data for selection of potential candidates. Seven proteins which absent in normal urine and blood, and could be secreted out by cancer cell lines were selected as candidates. Preliminarily wok of validation have been performed by western blot, IHC and ELISA in different cancer cell lines and urine samples from bladder cancer patients and normal individuals. We will further validate these candidates in more clinical samples. The results of validation in clinical samples will help us to discover the potential bladder cancer biomarkers in urine.