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J Proteome Res. 2016 Jul 1;15(7):2164-77. doi: 10.1021/acs.jproteome.6b00106. Epub 2016 Jun 14.

A Comprehensive Investigation toward the Indicative Proteins of Bladder Cancer in Urine: From Surveying Cell Secretomes to Verifying Urine Proteins.

Guo J1,2, Ren Y3, Hou G1,2, Wen B3, Xian F1,2, Chen Z1, Cui P1, Xie Y1,2, Zi J3, Lin L3, Wu S4, Li Z4, Wu L1,2, Lou X1,2, Liu S1,2,3.

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CAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences , Beijing, 100101, China.
University of Chinese Academy of Sciences , Beijing, 100049, China.
Proteomics Division, BGI-Shenzhen , Shenzhen, Guangdong 518083, China.
Shenzhen Second People's Hospital , Shenzhen, Guangdong 518028, China.


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 propose 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, a total of 700 proteins were found with 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 that the combination of CO3 and LDHB was more sensitive as the cancer indicator than other groups. The discovery of the bladder cancer indicators through our strategy has paved an avenue to further biomarker validation.


2DE; MRM; biomarker; bladder cancer; iTRAQ; urine

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