Background Bladder cancer (BC) is one of the most common malignant cancer of urinary system in the worldwide. The purpose of the present study was to analysis differentially expressed genes (DEGs), biological pathways and prognostic significance BC by bioinformatics analysis. Methods The gene expression dataset GSE7476 and the mRNA Seq sequencing data were downloaded respectively from GEO and TCGA. A total of 220 DEGs were obtained in BC. GO analysis and KEGG pathway analysis were performed for up- and down-regulated DEGs. Then, a protein-protein interaction (PPI) networks and module were constructed by Cytoscape software. Survival analysis of hub genes was performed. Results The result of GO analysis revealed that the up-regulated DEGs were enriched mainly in sister chromatid segregation, while the down-regulated DEGs were enriched mainly in muscle contraction. The result of KEGG pathway analysis showed that up-regulated DEGs were enriched mainly in cell cycle, while down-regulated DEGs enriched in IL-17 signaling pathway. 41 hub gene and 3 crucial modules were identified in the PPI network. 15 genes significantly associated with patient prognosis in BC were obtained by Kaplan-Meier analysis. Conclusions In summary, the present study identified hub genes, crucial pathways and provide possible the molecular targets and prognostic biomarkers for targeted therapy and prognostic assessment of BC.