Detection of Cytochrome P450 and Other Drug-Metabolizing Enzyme mRNAs in Peripheral Blood Mononuclear Cells Using DNA Arrays

• Published in: Drug metabolism and disposition Vol. 28; no. 8; pp. 987 - 993
• Main Authors: NGUYEN, L. T, RAMANATHAN, M, WEINSTOCK-GUTTMAN, B, DOLE, K, MILLER, C, PLANTER, M, PATRICK, K, BROWNSCHEIDLE, C, JACOBS, L. D
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Pharmacology and Experimental Therapeutics 01-08-2000
• Subjects: Adult; Analytical, structural and metabolic biochemistry; ATP-Binding Cassette, Sub-Family B, Member 1 - biosynthesis; Biological and medical sciences; Cytochrome P-450 Enzyme System - analysis; Cytochrome P-450 Enzyme System - biosynthesis; Cytochrome P-450 Enzyme System - genetics; DNA-Binding Proteins - biosynthesis; Enzymes and enzyme inhibitors; Feasibility Studies; Female; Fundamental and applied biological sciences. Psychology; Humans; In Vitro Techniques; Leukocytes, Mononuclear - enzymology; Leukocytes, Mononuclear - metabolism; Liver - enzymology; Liver - metabolism; Male; Multidrug Resistance-Associated Proteins; MutS Homolog 3 Protein; Oligonucleotide Array Sequence Analysis - methods; Oxidoreductases; Quality Control; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - analysis; RNA, Messenger - biosynthesis; Human; Multiple sclerosis; Enzyme; Cytochrome P450; Liver; Gene expression; In vitro; Blood cell; Mononuclear cell; Messenger RNA; Gene; Fluorescent labelling; Drug-metabolizing enzyme; Comparative study
• ISSN: 0090-9556; 1521-009X

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.