An association between cholelithiasis and prostate cancer has been reported sporadically in previous case-control and experimental studies, suggesting that cholesterol may play a promotional role in prostate cancer development and progression. However, this relationship remains poorly understood, and population evidence based on a strict study design is needed. The authors examined the history of cholelithiasis and the development of prostate cancer in the Ohsaki cohort followed from 1995 to 2003, in which 230 new cases of prostate cancer were ascertained among 22,458 Japanese men. Baseline information, including history of cholelithiasis, was collected using a self-administered questionnaire. The Cox proportional hazards regression model was used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs). The results showed that patients with a history of cholelithiasis had a higher risk of prostate cancer with a multivariate adjusted HR of 1.72 (95% CI: 1.12–2.66), and especially for advanced prostate cancer, the corresponding value was 2.29 (95% CI: 1.21–4.35). The associations were robust after adjustment for different potential confounders. This population-based prospective cohort study indicates that a history of cholelithiasis is associated with an increased incidence of prostate cancer.