Physical activity may reduce colon cancer risk, but the underlying mechanisms remain unclear. Relating physical activity to cancer risk in anatomic segments of the colon may advance our understanding of possible mechanisms. We conducted a prospective study of 59,369 Norwegian men and women who were followed up for cancer incidence and mortality. Cox proportional hazards models were used to estimate multivariably adjusted hazard ratios (HR) and 95% confidence intervals (95% CI). All statistical tests were two sided. During 17 years of follow-up, 736 colon cancers and 294 rectal cancers were diagnosed. Overall, we found an inverse association between recreational physical activity and colon cancer risk, but subsite analyses showed that the association was confined to cancer in the transverse and sigmoid colon. The adjusted HR, comparing people who reported high versus no physical activity, was 0.44 (95% CI, 0.25-0.78) for cancer in the transverse colon and 0.48 (95% CI, 0.31-0.75) for cancer in the sigmoid colon. The corresponding HR for cancer mortality was 0.33 (95% CI, 0.14-0.76) for the transverse colon and 0.29 (95% CI, 0.15-0.56) for the sigmoid colon. For rectal cancer, there was no association with physical activity in these data. In conclusion, the inverse association of recreational physical activity with cancer risk and mortality in the transverse and sigmoid segments of the colon may point at increased colon motility and reduced fecal transit time as possible underlying mechanisms.