To investigate the combined effect of Ca and Fe on colon carcinogenesis, cell proliferation, apoptosis and fecal bile acids, male Wistar rats were fed the diet containing 5 g Ca/kg (normal Ca) or 15 g Ca/kg (excessive Ca) with 45 mg Fe/kg (normal Fe) or 500 mg Fe/kg (excessive Fe) for 32 days, and given an injection of 1,2-dimethylhydrazine on day 4. Supplemental Ca reduced colonic aberrant crypt foci (ACF), especially in excess Fe group. Excessive Fe elevated the ACF, especially in the normal Ca diet. When the Ca intake was high, excessive Fe caused no influence on the ACF. Alteration of colonic ACF was associated with those of liver and serum Fe concentration. Also, colonic cell proliferation and concentration of deoxycholic acid (DCA) in fecal water-soluble fraction were reduced by supplementation of dietary Ca, but unaffected by that of dietary Fe. Supplementation of Ca and/or Fe elevated colonic cell apoptosis. The results suggest that dietary Ca markedly suppresses colon ACF in the Fe-overloaded rats through altering Fe status, and that supplemental Ca lowers colonic cell proliferation and fecal DCA in the water-soluble fraction and elevates colonic cell apoptosis irrespective of Fe status.