Experimental studies suggest that increased urination frequency may reduce bladder cancer risk if carcinogens are present in the urine. Only 2 small studies of the effect of increased urination frequency on bladder cancer risk in humans have been conducted with conflicting results. Our purpose was to evaluate the effect of urination frequency on risk of bladder cancer in a large, multicenter case-control study. We analyzed data based on interviews conducted with 884 patients with newly diagnosed, bladder cancer and 996 controls from 1998 to 2001 in Spain. We observed a consistent, inverse trend in risk with increasing nighttime voiding frequency in both men (p 5 0.0003) and women (p 5 0.07); voiding at least 2 times per night was associated with a significant, 40-50% risk reduction. The protective effect of nocturia was apparent among study participants with low, moderate and high water consumption. The risk associated with cigarette smoking was reduced by nocturia. Compared with nonsmokers who did not urinate at night, current smokers who did not urinate at night had an OR of 7.0 (95% CI 5 4.7-10.2), whereas those who voided at least twice per night had an OR of 3.3 (95% CI 5 1.9-5.8) (p value for trend 5 0.0005). Our findings suggest a strong protective effect of nocturia on bladder cancer risk, providing evidence in humans that bladder cancer risk is related to the contact time of the urothelium with carcinogens in urine. Increased urination frequency, coupled with possible dilution of the urine from increased water intake, may diminish the effect of urinary carcinogens on bladder cancer risk. ' 2008 Wiley-Liss, Inc.Key words: urination frequency; smoking; water intake; bladder neoplasms; epidemiology Experimental evidence suggests that if carcinogens are present in the urine, frequent urination may reduce bladder cancer risk. Three lines of limited evidence support this hypothesis. First, dogs exposed to the bladder carcinogen 2-napthylamine who have undergone surgical diversion of the urine do not develop tumors in the part of the bladder with no urine contact. 1 Second, dogs administered the bladder carcinogen 4-aminobiphenyl and catheterized to regulate urination frequency have decreased levels of urothelial DNA adducts with increased urination frequency. 2 Third, urine appears to be a promoter of bladder carcinogenesis in rat. 3,4 A protective effect of increased urinary frequency is further supported by the results of 2 observational studies. In high-risk areas for bladder cancer in Israel, infrequent micturition and high urine concentration were more prevalent than in low-risk areas. 5 In Serbia, a significant protective effect was associated with increased urination frequency based on 130 cases and 130 controls in a hospital-based case-control study. 6 This study had a number of limitations, however, including small sample size and minimal information on methods used to determine urination frequency was provided.Our purpose was to evaluate the effect of urination frequency on risk of urothelial (tran...