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Effects of Co-exposure of Di-(2-ethylhexyl)Phthalate and Benzo(a)pyrene on CYP1A1 and CYP3A4 Enzyme Activities in HepG2 Cells

[Objective] Effects of co-exposure of Di-(2-ethylhexyl)phthalate and Benzo(a)pyrene on CYP1A1 and CYP3A4 enzyme activities were investigated in HepG2 cells.[Methods] HepG2 cells were treated with either 64.0μmol/L B(a)P alone or DEHP alone(62.5,125.0,250.0,500.0,1000.0μmol/L) or DEHP at the indicated concentrations plus 64.0μmol/L B(a)P or dimethyl sulfoxide(DMSO,solvent control,1‰) for 48h and 72h.The cell proliferation was evaluated by CCK8 assay.The activities of CYP 1A-associated deethylation of ethoxyresorufin(EROD) and CYP3A-associated ethoxycoumarin-O-deethylation(ECOD) were measured.Expressions of CYP1A1 and CYP3A4 at mRNA and protein were detected by quantitative real time PCR method and Western blot,respectively.[Results] The cell viability was decreased significantly in all co-exposure of B(a)P and DEHP groups compared with the controls(P0.01).However,the EROD and ECOD activities significantly increased,when compared with the DEHP alone treated group(P0.05,P0.01).Only up-regualtion of CYP1A1 transcriptional levels were observed in all co-exposure of B(a)P and DEHP groups.However,the levels of mRNA and protein expression of CYP3A4 were increased(P0.01),when compared with the DEHP alone treated group.[Conclusion] Co-exposure of DEHP at certain concentrations and B(a)P(64.0μmol/L) induced the increases in the ERODandECODactivities,aswellastheCYP1A1transcriptional levelandtheCYP3A4transcriptionalandproteinlevels.