 | Melvili Cintron University of Puerto Rico, Mayagüez Industrial BiotechnologyMentor(s)A.-N. Tony Kong, Ph.D. Constance Saw Ying Huang Tien-Yuan Wu Department of Pharmaceutics Rutgers University |
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| Effects of dietary phytochemical cancer chemopreventive compounds in the transcriptional activation of the Nrf2-antioxidant signaling pathways leading to cancer prevention | ||
| Phytochemicals are health beneficial chemicals produced in plants. During this study we examined the effects of Indole-3-carbinol (I3C), found abundantly in cruciferous vegetables such as broccoli and Brussel sprouts, and its active metabolite 3,3-di-indolylmethane (DIM), on the antioxidant responsive element (ARE)-mediated signaling and gene expression in order to elucidate their potential cancer preventive effects. ARE is a regulatory element of many phase II drug-metabolizing/detoxification enzymes and cellular defensive enzymes regulated by nuclear factor erythroid-related factor 2 (Nrf2) transcription factor. Human liver cancer cell line (HepG2-C8) was transfected with ARE-luciferase gene and tested with 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay to determine the cytotoxicity of the compounds at different doses. Phenethyl isothiocyanate (PEITC) and sulforaphane (SFN) were used as positive controls, and DMSO 0.1% as an equivalent to no treatment control. ARE luciferase assay was used to quantify the amount of ARE-mediated induction being expressed after being stimulated by different doses of I3C, DIM, PEITC and SFN. The combination indexes of I3C with SFN or PEITC and DIM with SFN or PEITC in stimulating ARE-luciferase protein expression were also examined to test any potential synergistic effect between the combinations treatments. Other methods such as Western Blot and q-PCR to measure protein and mRNA expressions of Nrf2 and its downstream anti-oxidative stress enzyme, hemoxygenase-1 (HO-1), will also be conducted. |