In the present studies, we utilized prostate cancer cell culture models to elucidate the mechanisms of action of broccoli-derived phytochemicals 3,3′-diindolylmethane (DIM) and indole-3-carbinol (I3C). We found DIM and I3C at 1–5 µM inhibited androgen and estrogen-mediated pathways and induced xenobiotic metabolism pathway. By contrast, DIM and I3C induced cyclin inhibitors, indicators of stress/DNA damage, only at ≥25 µM. We also demonstrated that an inhibitory effect of DIM and I3C on cell growth involves inhibition of insulin-like growth factor-1 receptor expression. More importantly, we showed that differences in efficacies and mechanisms existed between DIM and I3C. These included differences in effective concentrations, a differential effect on androgen receptor binding, and a differential effect on xenobiotic metabolic pathway through aryl hydrocarbon receptor-dependent and -independent mechanism. Furthermore we determined that several other diet-derived cancer protective compounds, similar to DIM and I3C, exhibited pleiotrophic effects on signaling pathways that included proliferation, cell cycle, and nuclear receptors-mediated pathways. However, the efficacies and mechanisms of these compounds vary. We also showed that some cellular pathways are not likely to be affected by DIM or I3C when circulating concentration of orally ingested DIM or I3C is considered. Based on our results, a model for cancer protective effects of DIM and I3C was proposed. © 2011 Wiley Periodicals, Inc.