Researches had proven that the occurrence of bladder cancer (BC) is much higher in men than those in women, which induced us to explore whether androgen plays a role in BC. A total of 147 patients who were diagnosed with primary BC by histopathological biopsy were included. Meanwhile 154 non-tumor patients were matched as the control group. The continuous variables were expressed as median (interquartile range, IQR) and compared by Mann–Whitney U test, for the reason that the data were not matched the requirementsthe of normal test. A Chi-square test was used to compare the categorical variables, which were expressed as frequency (percentage). Meanwhile univariate and multivariate logistic regression was done to further evaluating the potential independent factor of BC. P < 0.05 was considered statistically significant. Univariate multivariate analyse showed significant difference between two groups in hemoglobin (OR 0.979, 95% CI 0.968–0.991, P < 0.001), hypertension (OR 3.026, 95% CI 1.731–5.288, P < 0.001), diabetes (OR 4.294, 95% CI 1.887–9.771, P = 0.001) and smoking (OR 1.729, 95% CI 1.096–2.729, P = 0.019). Furthermore, multivariate logistic regression analysis was conducted to eliminate the interference of confounding factors, which showed that testosterone seems to be great correlated with the BC (OR 1.002, 95% CI 1.000–1.003, P = 0.017). Similar results were also found in hemoglobin (OR 0.981, 95% CI 0.968–0.993, P = 0.002), hypertension (OR 2.780, 95% CI 1.509–5.120, P = 0.001), diabetes (OR 3.313 95% CI 1.373–7.991, P = 0.008) and smoking (OR 1.938, 95% CI 1.184–3.174, P = 0.009). As a conclusion, our study showed that there was significant correlation between serum total testosterone levels and the occurrence of BC, similar results were shown in hemoglobin, hypertension, diabetes and smoking.