The Estrogen Metabolites (2-hydroxyestrogens: 16α-hydroxyestrone) Urine Ratio (EMUR) has been negatively associated with breast cancer; Mexican women have a lower EMUR than other populations. We evaluated the effectiveness of 3,3'-diindolylmethane (DIM) supplementation on increasing EMUR in premenopausal women. A randomized, double-blind clinical trial (NCT02525159 at ClinicalTrial.gov) was carried out on 60 women with an EMUR below 0.9. Patients were assigned randomly to receive a placebo or 75 mg of DIM a day (administered as 300 mg of DIM-BR®) for 30 day. Urine samples were obtained at baseline, at 30 day of supplementation, and 30 day after finishing supplementation. A Mann-Whitney U test was used to compare the EMUR; an ANOVA was used to analyze differences in body composition. EMUR was analyzed using ESTRAMET™ kits. While DIM-treated subjects did not increase their EMUR at 30 day of supplementation (p > 0.05), there was a non-significant positive trend 30 day once supplementation ended (p = 0.06). The DIM group saw a more significant decrease in body fat percentage than the placebo group (p = 0.04). In premenopausal Mexican women, 75 mg of the daily DIM supplement was ineffective in increasing EMUR; further studies are needed to evaluate the effective dosage, time frames, and effect on body fat.