Study On The Early Biomarkers For PAH Exposure In Mammalian Cell

Polycyclic aromatic hydrocarbons (PAHs) are environmentally prevalent carcinogens found in combustion products such as cigarette smoke and diesel exhaust. It is implicated in tumor initiation, promotion and progression .Benzo[a]pyrene (B[a]P),one of the most common and extensively PAHs studied, exhibits strong carcinogenic properties in several animal species including human . In vivo, B[a]P is mainly metabolized into 7,8-dihydro-diol-9, 10-epoxide benzo(a) pryrene (BPDE), which can eventually induce adduct formation or oxidative damage to DNA followed by mutagenesis and tumorigenesis, Many studies have focused on understanding B[a]P and its metabolite—BPDE induced mutagenesis and carcinogenesis.We here use the proteomics method to concentrate on investigating the differences of extracellular protein profile of FL cells treated by BPDE. It induces comprehensivechanges in the protein expression profile of human amnion FL cells, including the induction, suppression of various proteins. The experiments were carried that FL cells were exposed to 5umol/L BPDE for 2. 5h, as same control FL cells cultures was treated by the solvent, dimethylsulfoxide (DMSO). After the chemicals were removed completely, the cells were incubated in fresh medium for 12h and another 6h followed by the serum-free media. The serum-free media from cultured FL cells were concentrated by ultrafiltration and the concentrated extracellular proteins were separated by two-dimensional polyacrylamide gel electrophoresis and visualized by silver staining;the gels were digitized prior to computer-based matching and quantitative analysis with image analysis software. The digitized images then were analyzed with 2D analysis software in order to establish the differential protein profile of FL cells after BPDE treatment based on the 2-DE mapping. The profiles wereanalyzed with image analysis software. We found that 4 protein spots were induced by BPDE and 3 protein spots were repressed. We conclude that there are alteration in the extracellular protein profile of BPDE treated FL cells. Therefore these proteins may help better understand the cellular stress responses to chemical carcinogens, and may lead to new studies on the functions of these BPDE-responsive proteins. Furthermore, some of these proteins may serve as candidates of biomarker for detecting exposure of human populations to environmental carcinogens.