title: Single nucleotide polymorphism of CYP3A4 intron 2 and its influence on CYP3A4 mRNA expression and liver enzymatic activity in human liver.
abstract: In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of endogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism (SNP) of CYP3A4 intron 2 in the liver and its effects on the mRNA expression and enzymatic activity of CYP3A4, genomic DNA was extracted from 96 liver tissue samples obtained from patients who had undergone liver surgery. An SNP of CYP3A4 intron 2 was identified by polymerase chain reaction (PCR)-single-strand confirmation polymorphism and DNA sequencing. The mRNA expression of CYP3A4 was determined by the fluorescence quantitative PCR technique. The enzymatic activity of CYP3A4 was measured using erythromycin and testosterone as probe substrates. Twelve patients were found to have the SNP/T4127G CYP3A4 within intron 2. The mRNA levels of CYP3A4 in wild-type and SNP/T4127G samples were 2.62±1.09 and 2.79±1.63, respectively (P>0.05). Erythromycin N-demethylase activity in wild-type and SNP/T4127G samples were 121.2±32.8 and 124.7±61.6 nmol·mg(-1)·min(-1), respectively (P>0.05). The activity of testosterone 6β-hydroxylase was significantly different between wild-type (648±173 pmol·mg(-1)·min(-1)) and SNP/T4127G samples (540±196 pmol·mg(-1)·min(-1); P<0.05). In conclusion, the SNP/T4127G of CYP3A4 intron 2 exists in the liver. This SNP does not affect the mRNA expression of CYP3A4 but significantly decreases the hepatic microsomal testosterone 6β-hydroxylase activity of CYP3A4. Furthermore, this study indicates that the appropriate selection of probe substrates is very important in studying the relationship between the genotype and phenotype of CYP3A4.
authors.affiliation: Department of Endocrinology, Hubei Integrative Chinese and Western Medicine Hospital, Wuhan, 430015, China.|Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan, 430071, China.
authors.forename: Min
authors.initials: M
authors.lastname: Huang
authors.affiliation: Department of Endocrinology, Hubei Integrative Chinese and Western Medicine Hospital, Wuhan, 430015, China.
authors.forename: Han-Ming
authors.initials: HM
authors.lastname: Wang
authors.affiliation: Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan, 430071, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan, 430071, China.|Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan, 430071, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Ping
authors.affiliation: Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan, 430071, China.
authors.forename: Man
authors.initials: M
authors.lastname: Chen
authors.affiliation: Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan, 430071, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Xu
authors.affiliation: Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan, 430071, China. wanghui19@whu.edu.cn.|Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan, 430071, China. wanghui19@whu.edu.cn.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
mesh_terms: D000328:Adult; D044466:Asian People / Q000235:genetics*; D002681:China; D051544:Cytochrome P-450 CYP3A / Q000235:genetics* / Q000378:metabolism*; D004917:Erythromycin / Q000378:metabolism; D005838:Genotype; D006801:Humans; D007438:Introns; D008099:Liver / Q000201:enzymology*; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D012333:RNA, Messenger / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods; D013739:Testosterone / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D013739:Testosterone; D004917:Erythromycin; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human