Effects of zearalenone on mRNA expression and activity of cytochrome P450 1A1 and 1B1 in MCF-7 cells

Abstract

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous pollutant and promoter of carcinogenesis with both estrogenic and antiestrogenic effects in mammal epithelium. Zearalenone (ZEA) is a naturally occurring estrogenic contaminant of moldy feeds and is present in high concentrations in dairy products and cereals. Numerous studies describe a possible correlation between xenoestrogens and breast cancer risk. A potential mechanism for the etiology of breast cancer involves altered cytochrome P450 (CYP) enzymes. Since cocontamination of multiple compounds in our environmental and occupational circumstances likely happens and since few studies have addressed the molecular consequences of combinations of contaminants, we decided to investigate the effects of ZEA on basal and TCDD-induced mRNA expression and enzymic activity of CYP1A1 and CYP1B1 in human breast cancer MCF-7 cells. CYP1A1 enzyme activity was measured by the CYP1A1-referential activity assay, ethoxyresorufin O-deethylase (EROD), in MCF-7 cells. To investigate CYP1B1 activity, we employed the microsomal EROD assay prepared from baculovirus-infected insect cells expressing human cDNA CYP1B1. Reverse transcription-polymerase chain reaction was used to detect mRNA expression of CYP1A1 and CYP1B1 in MCF-7 cells. The results demonstrated that 10 nM TCDD could readily induce a significant increase in the enzyme activity and mRNA expression of CYP1A1 in MCF-7 cells and 5 nM estradiol (E2) significantly reduced both basal and TCDD-induced activity and mRNA expression in MCF-7 cells. The same pattern was observed with 50 nM ZEA. The estrogen receptor antagonist 4-hydroxytamoxifen could attenuate these inhibitive effects of both E2 and ZEA. Interestingly, Both E2 and ZEA could promote basal and TCDD-induced CYP1B1 activity but with no effect on CYP1B1 mRNA expression. These results suggest that the effect of ZEA on the TCDD-induced CYP1A1 activity and gene expression involved the estrogen receptor pathway and that the increase in the CYP1B1/CYP1A1 ratio underlying the basal or TCDD-treated condition might constitute one of the mechanisms underlying the synergic carcinogenic action of these compounds.