title: Detection of cytochrome P450 and other drug-metabolizing enzyme mRNAs in peripheral blood mononuclear cells using DNA arrays.
abstract: The purpose of this study was to determine whether the expression of cytochrome P450 (CYP) enzyme mRNAs, other drug-metabolizing enzyme mRNAs, and transporter mRNAs can be detected using DNA arrays. Total RNA was isolated from peripheral blood mononuclear cells of 10 multiple sclerosis patients and 10 age- and sex-matched controls. The mRNA was reverse transcribed to radiolabeled cDNA, and the resultant cDNA was used to probe a DNA array containing several thousand known human genes. The signals corresponding to several CYPs, drug-metabolizing, and transporter mRNAs was substantially above background. The results demonstrate that the DNA array technique has the sensitivity and the selectivity for applications in the pharmaceutical sciences. The mean values for mRNAs of specific CYPs and drug-metabolizing enzymes in peripheral blood cells were compared with reported values for liver. The capabilities of DNA arrays may prove useful for characterizing CYP expression in a variety of clinical samples.
authors.affiliation: Department of Pharmaceutics, State University of New York at Buffalo, Buffalo, New York, USA.
authors.forename: L T
authors.initials: LT
authors.lastname: Nguyen
authors.forename: M
authors.initials: M
authors.lastname: Ramanathan
authors.forename: B
authors.initials: B
authors.lastname: Weinstock-Guttman
authors.forename: K
authors.initials: K
authors.lastname: Dole
authors.forename: C
authors.initials: C
authors.lastname: Miller
authors.forename: M
authors.initials: M
authors.lastname: Planter
authors.forename: K
authors.initials: K
authors.lastname: Patrick
authors.forename: C
authors.initials: C
authors.lastname: Brownscheidle
authors.forename: L D
authors.initials: LD
authors.lastname: Jacobs
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis; D000328:Adult; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D005240:Feasibility Studies; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007963:Leukocytes, Mononuclear / Q000201:enzymology* / Q000378:metabolism; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D008297:Male; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D000074081:MutS Homolog 3 Protein; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011786:Quality Control; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D003577:Cytochrome P-450 Enzyme System; D004268:DNA-Binding Proteins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D000074081:MutS Homolog 3 Protein; D012333:RNA, Messenger; C078444:MSH3 protein, human; C421109:multidrug resistance-associated protein 1