The potential of soy isoflavones (SIs) to reduce colon cancer has been investigated in animal models. These studies have found that outcomes are variable and depend on SI dose. The present study investigated dose-response effects of SIs on colon carcinogenesis in a chemically induced rat cancer model. Sprague-Dawley male rats were injected with 1,2-dimethylhydrazine (DMH) and were provided experimental diets that contained 0, 10, 50, 150, or 500 mg of SI aglycones/kg of diet for 12 weeks. Plasma concentrations of genistein, daidzein, and equol were determined using time-resolved fluoroimmunoassay. Plasma concentrations of these SIs tended to increase in a dose-dependent manner in DMH-treated rats. The numbers of aberrant crypt foci (ACF) and the expression of cyclooxygenase-2 (COX-2) proteins of colons were significantly decreased in the SI-fed groups compared with the control group; however, suppression was not dose-dependent. Furthermore, there were no significant correlations between plasma SI concentrations and ACF or COX-2 expression. Increased SI intake and increased plasma levels of SIs and metabolites were not associated with tissue levels of lipid peroxidation. We conclude that dietary supplementation of SIs suppresses DMH-induced ACF formation and COX-2 expression in a dose-independent manner.