Screening Differentially Expressed Genes In Human Bladder Transitional Cell Carcinoma By CDNA Microarry And Verification

Objective To detect the gene expression profile of bladder transitional cell carcinoma (BTCC) and normal bladder mucous membrane tissues by high throughput cDNA microarray, screening differentially expressed genes, preliminary discussion on the origin and development of BTCC molecular mechanism, and to explore potential tumor specific gene.Methods1. To apply high throughput cDNA microarray to detecting6cases of BTCC and3cases of normal bladder mucous membrane tissues, screening differentially expressed genes, and then import them into CapitalBio the biomolecular functional annotation system MAS3.0. Get bioinformatics analysis incuding GO function classification and pathway.2. Using real-time quantitative RT-PCR to detect four significant differences in gene expression, and validate the reliability of the result of the gene chip.Results1. The chip results were screened263bladder transitional cell carcinoma common differentially expressed genes. There are92genes down-regulation more than2times and171genes down-regulation more than2times of gene expression. Differences in gene GO function classification involved DNA transcription and transcriptional regulation, cell division, cell cycle, decomposition of protein metabolism, apoptosis, and resistance to apoptosis, cell proliferation, immune response, DNA replication and repair, etc. Differences in gene pathway involved in cell cycle, DNA polymerization pathway, ubiquitin protein pathways mediating, mismatch repair pathway, P53signaling pathway, oxidative phosphorylation pathway, cell factor and its receptor interaction pathways, PPAR signaling pathways, TGF-beta signaling pathways, etc.2. Real-time quantitative RT-PCR, according to the results of four genes TOP2A, CDK1, BIRC5A, the relative expression CAV1were10.45,5.35,28.46,0.14, and the overall trend of gene chip results are basically identical.Conclusion 1. Gene chip screening of differentially expressed genes, GO function enrichment and pathway enrichment analysis, fully proving the occurrence of BTCC is a more complex of gene regulation, multi-step, multi-channel process. This experiment screening differentially expressed genes and tumor related genes is of great importance to the diagnosis of bladder cancer.2. Real-time fluorescent quantitative PCR test TOP2A, CDK1, B1RC5A, CAV1gene expression in chip results, further verify the reliability of chip results, and found that these genes may be the most the potential biomarkers in the diagnosis of BTCC.