In vitro and in vivo studies have shown that carotenoid supplementation is associated with decreased DNA damage, but the role of dietary carotenoids in cancer risk remains controversial because epidemiologic studies have yielded conflicting results. Limited data exist regarding the role of dietary carotenoids in the context of constitutional genetic instability in cancer risk. This case-control study estimated dietary carotenoid intake [microg/(kJ . d)] from a FFQ for 423 patients with bladder cancer and 467 healthy controls, and quantified baseline and benzo[a]pyrene diol epoxide (BPDE)- and gamma-radiation-induced DNA damage in the peripheral blood lymphocytes using the comet assay. Overall, intake of total carotenoids was lower (P < 0.01) for bladder cancer cases (mean +/- SD: 1273.4 +/- 688.9) compared with healthy controls (1501.3 +/- 791.5). When categorized into quartiles, there was an inverse association between increasing levels of carotenoid intake and bladder cancer risk with greatest protective effect [odds ratio (OR) = 0.56, 95% CI, 0.37-0.85] in the quartile with the highest level of intake. Baseline and mutagen-induced DNA damage was significantly higher in cases than in controls; when analyzed jointly with carotenoid intake, high DNA damage and low carotenoid intake were associated with the highest risk. For example, with high baseline DNA damage and low total carotenoid intake, the OR was 3.08 (95% CI, 1.64-5.77); with high baseline DNA damage and high total carotenoid intake, the risk was somewhat attenuated (OR = 2.49, 95% CI, 1.28-4.84). The risk was decreased further for low baseline DNA damage and low total carotenoid intake (OR = 2.18; 95% CI, 1.13-4.22). This study provides evidence of a preventive role for carotenoids in bladder cancer, and these data may have important implications for cancer prevention, especially for individuals susceptible to DNA damage.