Abstract Background: Acrylamide has been classified as a probable human carcinogen based chiefly on laboratory evidence. However, the influence of dietary acrylamide intake on risk of esophageal, gastric, and colorectal cancer has not been extensively studied. We aimed to evaluate the association between dietary acrylamide intake and esophageal, gastric, and colorectal cancer using data from the Japan Public Health Center–based Prospective Study. Methods: Our study included 87,628 participants who completed a food-frequency questionnaire at enrollment in 1990 and 1993. We used Cox proportional hazards regression models to estimate hazards ratios and 95% confidence intervals (CI) after adjusting for confounding factors. Results: After 15.5, 15.3, and 15.3 mean years of follow-up for esophageal, gastric, and colorectal cancer, we identified and analyzed 391 esophageal, 2,218 gastric, and 2,470 colorectal cancer cases, respectively. Compared with the lowest quintile of acrylamide intake, the multivariate HR for the highest quintile was 0.86 (95% CI, 0.53–1.39; Ptrend = 0.814), 0.84 (95% CI, 0.69–1.01; Ptrend = 0.301), and 0.93 (95% CI, 0.79–1.08; Ptrend = 0.165) for esophageal, gastric, and colorectal cancer, respectively, in the multivariable-adjusted model. Furthermore, no significant associations were observed when the participants were stratified by cancer subsites. Conclusions: In conclusion, this study demonstrated that dietary acrylamide intake was not associated with increased risk of esophageal, gastric, or colorectal cancer among the Japanese population. Impact: It is the first time to assess the effect of dietary acrylamide intake on risk of digestive system cancer in Asian populations.