Urine is an ideal material to study the cancer-related protein biomarkers in bladder, whereas exploration to these candidates is confronting technique challenges. Herein, we proposed a comprehensive strategy of searching the urine proteins related with bladder cancer. The strategy consists of three core combinations, screening the biomarker candidates in the secreted proteins derived from the bladder cancer cell lines and verifying them in patient urines, defining the differential proteins through two-dimensional electrophoresis (2DE) and isobaric tags for relative and absolute quantitation (iTRAQ) coupled with LC MS/MS, and implementing quantitative proteomics of profiling and targeting analysis. With proteomic survey, total of 700 proteins were found their abundance of secreted proteins in cancer cell lines different from normal, while 87 proteins were identified in the urine samples. The multiple reaction monitoring （MRM）-based quantification was adapted in verifying the bladder cancer related proteins in individual urine samples, resulting in 10 differential urine proteins linked with the cancer. Of these candidates, receiver operating characteristic analysis revealed the combination of CO3 and LDHB was more sensitive as the cancer indication than other groups. The discovery of the bladder cancer indicators through our strategy has paved an avenue to further biomarker validation.
Journal of proteome research. 2016 Jun 04 [Epub ahead of print]
Jiao Guo, Yan Ren, Guixue Hou, Bo Wen, Feng Xian, Zhen Chen, Ping Cui, Yingying Xie, Jin Zi, Liang Lin, Song Wu, Zesong Li, Lin Wu, Xiaomin Lou, Siqi Liu