Background: We aimed to explore the expression of forkhead box class O (FOXO) and relations between expressions of FOXOs and clinicopathological characteristics and prognosis of bladder cancer. Methods: We enrolled a cohort of 276 patients with bladder cancer in our study. Expressions of FOXOs in bladder cancer tissue and adjacent tissue were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC). Correlations between expression of FOXOs and clinicopathological characteristics and prognosis were analyzed. The relationship between expression of FOXOs and survival time of patients with bladder cancer was analyzed by the Kaplan-Meier survival analysis and the Log-rank test; individual variables which may affect the prognosis of bladder cancer were detected by the Cox proportional hazard regression model. Results: Compared with bladder cancer tissue, a higher expression of FOXOs was detected in paracancerous tissue. We found significant associations between histological grade and the expressions of FOXOs, clinical stage and the expressions of FOXOs, and lymph node metastasis and the expressions of FOXOs (all P < 0.05). When used for diagnosing bladder cancer, the mRNA expression of FOXO1/3/4 produced cut off values of 1.475, 1.305, and 1.295, respectively, exhibiting relatively high specificity and sensitivity. The Kaplan-Meier curves indicated that patients with a higher expression of FOXOs tended to have a longer overall survival than those with lower expression. The Cox regression analysis revealed that lymph node metastasis, high clinical stage, and low expression of FOXOs were independent risk factors for bladder cancer prognosis. Conclusion: Our results indicate that the expression of FOXOs is closely correlated with clinicopathological characteristics and prognosis of bladder cancer.