RESUMO
Exposure to environmental toxicants or exogenous oestrogen increases the risk of cancer. Some toxicants such as polycyclic aromatic hydrocarbons (PAH) undergo biotransformation to become genotoxic agents. Cytochrome p450 (CYP) 1B1 is an enzyme catalysing this transformation. Consumption of fruit and vegetables is considered to be protective against carcinogenesis, and naringenin can be found abundantly in citrus fruits. In the present study, the effect of naringenin on the regulation of CYP1B1 was investigated in MCF-7 cells. Enzyme inhibition assays revealed that naringenin inhibited CYP1B1 at or above 5 µm but not CYP1A1 activity. Quantitative PCR analysis also demonstrated that 1 µm-naringenin reduced CYP1B1 mRNA expression induced by 7,12-dimethylbenz(α)anthracene (DMBA). Further study illustrated that the suppression was at the transcriptional level. Since previous studies have shown that oestrogen response element (ERE) and xenobiotic-responsive element (XRE) are functional binding sequences in the promoter region of CYP1B1, interference of DNA binding on these two elements was pursued. Employing reporter gene assays as well as the electromobility shift assay, we verified that naringenin counteracted DMBA-induced XRE binding at − 1675. These results supported the notion that fruit consumption could be a protective measure against PAH biotransformation.
Assuntos
Hidrocarboneto de Aril Hidroxilases/genética , Flavanonas/farmacologia , Ativação Transcricional/efeitos dos fármacos , Xenobióticos/metabolismo , 9,10-Dimetil-1,2-benzantraceno/metabolismo , Sítios de Ligação , Linhagem Celular Tumoral , Citocromo P-450 CYP1B1 , Ensaio de Desvio de Mobilidade Eletroforética , Genes Reporter , Humanos , Luciferases/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
Polycyclic aromatic hydrocarbons (PAH) are established cancer initiators that can be found in our food and environment. Some dietary phytochemicals are strong inhibitors of PAH-induced mutagenesis. The soya isoflavone genistein has been shown previously in our laboratory to be an inhibitor of PAH metabolite binding to DNA. In the present study, we investigated the effect of genistein on oxidative DNA damage induced by PAH in the non-tumorigenic breast cell line MCF10A. 7,12-Dimethyl-benz[a]anthracene (DMBA) can induce expressions of CYP1A1 and CYP1B1 which are known to be responsive to PAH. These enzymes, in turn, will metabolise the PAH into their ultimate carcinogenic forms. Genistein can significantly suppress the expressions within 5 microm. The comet assay indicated that DMBA introduced DNA damage to these cells, and co-treatment with genistein at 5 or 10 microm could alleviate the damage. In addition to the chelation of DMBA metabolites to DNA, flow cytometry results revealed that oxidation was also a factor of DNA damage. The oxidative DNA damage could be removed by co-treating with 10 microm-genistein. Because no increased oxidative DNA repair was observed, suppression on the cytochrome enzymes appeared to be the underlying mechanism.
Assuntos
Poluentes Atmosféricos/metabolismo , Anticarcinógenos/farmacologia , Mama/metabolismo , Dano ao DNA , Genisteína/farmacologia , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , 9,10-Dimetil-1,2-benzantraceno , Hidrocarboneto de Aril Hidroxilases/genética , Mama/citologia , Mama/efeitos dos fármacos , Linhagem Celular , Ensaio Cometa , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1B1 , Feminino , Citometria de Fluxo , Expressão Gênica/efeitos dos fármacos , Humanos , Estresse Oxidativo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodosRESUMO
Licorice is derived from the rhizomes of Glycyrrhiza glabra. It has been used for confectioneries or culinary purposes. The rhizomes contain many flavonoidal compounds that have been shown to be biologically active. In the present study, effect of the licorice flavonoid isoliquiritigenin (ILN) on polycyclic aromatic hydrocarbon (PAH)-induced XRE transactivation and the downstream expression were investigated in MCF-7 cells. The environmental toxicant PAHs are pro-carcinogens and are biotransformed into their ultimate genotoxic structures by cytochrome P450 (CYP) 1 enzymes. Reporter gene assay revealed that ILN reduced XRE transactivation triggered by 7,12-dimethylbenz[α]anthracene (DMBA) or 2,3,7,8-Tetrachlorodibenzodioxin (TCDD). Our EMSA results also demonstrated that the flavonoid diminished DMBA-induced XRE binding. The reduced transactivation could be the result of a decreased amount of AhR translocating from cytosol to nucleus as shown in Western analysis. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay demonstrated that expressions of genes with XRE-containing promoters, including CYP1A1, 1A2, and 1B1, followed the same pattern of XRE transactivation. The present study illustrated that ILN might downregulate PAH-induced expressions through antagonizing AhR translocation.