Two-dimensional electrophoresis for the screening of biomarker in carcinomas of the urinary bladder / 中华检验医学杂志

ABSTRACT

Objective This study was designed to establish and optimize the two-dimensional gel electrophoresis (2-DE) for the proteome analysis of human Transitional Cell Carcinomas of the Urinary Bladder ,and to established the two-dimensional gel electrophoresis (2-DE) profile of human Transitional Cell Carcinomas of the Urinary Bladder(TCC) of different stages in carcinogenic process. The basic aim is to perform differential analysis and provide a basis for identifying carcinogenesis-associated protein of TCC.Methods After obtaining samples of the normal,invasive tissue,noninvasive tissue,a series of methods Such as sample preparation,electrophoresis parameters,gel concentration,PDQuest 2DE analysis software,peptide mass fingerprint (PMF,MALDI-TOF-MS)were used to analyses these tissues.Results The good 2DE patterns of different tissues including resolution and reproducibility were obtained. After coomassie Brilliant Blue R-250 staining, the average matching ratio was 76%.There was a good reproducibility of spot position in 2-DE map, with average deviation in IEF direction of (0.98?0.26) mm, while in SDS-PAGE direction it was(1.23?0.22) mm. The 2-DE patterns with good quality had been obtained. Ten protein spots chosen randomly from different stages were identified by PMF, some of which were involved in the cell proliferation, differentiation, cycle regulation, and tumor occurrence, such as Heat shock protein-27,SFN etc.Conclusions The results provide a fundamental basis for further study of mechanism of human Transitional Cell Carcinomas of the Urinary Bladder and screening its specific marker.