Identification of miRNAs that regulate human CYP2B6 expression

Human hepatic cytochrome P450 2B6 (CYP2B6) expressed is responsible for the metabolism of many drugs, such as cyclophosphamide, ifosfamid, and efavirenz. In the present study, the correlation between CYP2B6 mRNA and protein levels in human liver samples was found to be moderate, indicating a contribution of posttranscriptional regulation of CYP2B6. Thus, we examined the role of microRNAs (miRNAs) in the regulation of CYP2B6. We established two kinds of HEK293 cell lines stably expressing CYP2B6, including or excluding the full-length 3′-untranslated region (3′-UTR) (HEK/2B6+UTR and HEK/2B6 cells, respectively). We tested 14 miRNAs that were predicted to bind to the 3′-UTR of CYP2B6 and found that the overexpression of miR-145, miR-194, miR-222, and miR-378 decreased the CYP2B6 protein level and activity in HEK/2B6+UTR but not in HEK/2B6 cells. These results suggested that miR-145, miR-194, miR-222, and miR-378 negatively regulate CYP2B6 expression by binding to the 3′-UTR. A negative correlation was not observed between the translational efficiency of CYP2B6 and the expression level of miR-145, miR-194, miR-222, or miR-378. This is due to the contribution of multiple miRNAs to CYP2B6 regulation. In conclusion, this study demonstrated that human CYP2B6 is posttranscriptionally regulated by miR-145, miR-194, miR-222, and miR-378.