title: Single-Nucleotide Polymorphisms in Cytochrome P450 2E1 (CYP2E1) 3'-Untranslated Region Affect the Regulation of CYP2E1 by miR-570.
abstract: Human cytochrome P450 2E1 (CYP2E1) catalyzes the metabolism of numerous xenobiotics, including acetaminophen and ethanol. CYP2E1 expression is known to be extensively regulated by post-transcriptional and post-translational mechanisms. A previous study had reported that a single-nucleotide polymorphism (SNP) 1561A>G in the 3'-untranslated region (3'-UTR) of CYP2E1 leads to a decreased CYP2E1 mRNA level in human peripheral blood mononuclear cells. In this study, we examined the possibility that microRNA(s) (miR) may be involved in the SNP-mediated modulation of CYP2E1 expression. Genotyping and sequencing analyses revealed that another SNP, 1556T>A, in the 3'-UTR was in complete linkage disequilibrium with the SNP 1561A>G. We termed the alleles with 1556T and 1561A or 1556A and 1561G haplotype I or II, respectively. A luciferase assay revealed that miR-570 recognizes the CYP2E1 3'-UTR of haplotype I but not haplotype II. Human embryonic kidney 293 (HEK293) cell lines stably expressing human CYP2E1 that included the 3'-UTR of haplotype I or II (HEK/2E1(I) or HEK/2E1(II) cells, respectively) were established. Overexpression of miR-570 significantly decreased the CYP2E1 protein level in the HEK/2E1(I) cells but not in the HEK/2E1(II) cells. In seven human livers with diplotype I/I, the CYP2E1 protein levels were inversely correlated with the miR-570 levels, but no relationship was observed in 25 human livers with diplotypes I/II and II/II. Collectively, it was demonstrated that human CYP2E1 was regulated by miR-570 in a genotype-dependent manner. This report describes the first proof that SNP in 3'-UTR of human P450 affects binding of miRNA to modulate the expression in the liver.
authors.affiliation: Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa, Japan (Ma.N., T.M., T.F., Mi.N.); Department of Legal Medicine, School of Medicine, Iwate Medical University, Morioka, Japan (M.T., Y.A.); Department of Medicine, School of Medicine, Washington University, St. Louis, Missouri (H.L.M.).
authors.forename: Masataka
authors.initials: M
authors.lastname: Nakano
authors.affiliation: Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa, Japan (Ma.N., T.M., T.F., Mi.N.); Department of Legal Medicine, School of Medicine, Iwate Medical University, Morioka, Japan (M.T., Y.A.); Department of Medicine, School of Medicine, Washington University, St. Louis, Missouri (H.L.M.).
authors.forename: Takuya
authors.initials: T
authors.lastname: Mohri
authors.affiliation: Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa, Japan (Ma.N., T.M., T.F., Mi.N.); Department of Legal Medicine, School of Medicine, Iwate Medical University, Morioka, Japan (M.T., Y.A.); Department of Medicine, School of Medicine, Washington University, St. Louis, Missouri (H.L.M.).
authors.forename: Tatsuki
authors.initials: T
authors.lastname: Fukami
authors.affiliation: Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa, Japan (Ma.N., T.M., T.F., Mi.N.); Department of Legal Medicine, School of Medicine, Iwate Medical University, Morioka, Japan (M.T., Y.A.); Department of Medicine, School of Medicine, Washington University, St. Louis, Missouri (H.L.M.).
authors.forename: Masataka
authors.initials: M
authors.lastname: Takamiya
authors.affiliation: Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa, Japan (Ma.N., T.M., T.F., Mi.N.); Department of Legal Medicine, School of Medicine, Iwate Medical University, Morioka, Japan (M.T., Y.A.); Department of Medicine, School of Medicine, Washington University, St. Louis, Missouri (H.L.M.).
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Aoki
authors.affiliation: Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa, Japan (Ma.N., T.M., T.F., Mi.N.); Department of Legal Medicine, School of Medicine, Iwate Medical University, Morioka, Japan (M.T., Y.A.); Department of Medicine, School of Medicine, Washington University, St. Louis, Missouri (H.L.M.).
authors.forename: Howard L
authors.initials: HL
authors.lastname: McLeod
authors.affiliation: Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa, Japan (Ma.N., T.M., T.F., Mi.N.); Department of Legal Medicine, School of Medicine, Iwate Medical University, Morioka, Japan (M.T., Y.A.); Department of Medicine, School of Medicine, Washington University, St. Louis, Missouri (H.L.M.) nmiki@p.kanazawa-u.ac.jp.
authors.forename: Miki
authors.initials: M
authors.lastname: Nakajima
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics*; D019392:Cytochrome P-450 CYP2E1 / Q000235:genetics* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008862:Microsomes, Liver / Q000502:physiology; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011485:Protein Binding / Q000502:physiology
chemical_list: D020413:3' Untranslated Regions; C582115:MIRN570 microRNA, human; D035683:MicroRNAs; D019392:Cytochrome P-450 CYP2E1