Characterization of increased drug metabolism activity in dimethyl sulfoxide (DMSO)-treated Huh7 hepatoma cells.
The objective of this study was to characterize Huh7 cells' baseline capacity to metabolize drugs and to investigate whether the drug metabolism was enhanced upon treatment with dimethyl sulfoxide (DMSO). The messenger RNA (mRNA) levels of major Phase I and Phase II enzymes were determined by quantitative real-time-polymerase chain reaction (RT-PCR), and activities of major drug-metabolizing enzymes were examined using probe drugs by analysing relevant metabolite production rates. The expression levels of drug-metabolizing enzymes in control Huh7 cells were generally very low, but DMSO treatment dramatically increased the mRNA levels of most drug-metabolizing enzymes as well as other liver-specific proteins. Importantly, functionality assays confirmed concomitant increases in drug-metabolizing enzyme activity. Additionally, treatment of the Huh7 cells with 3-methylcholanthrene induced cytochrome P450 (CYP) 1A1 expression. The results indicate that DMSO treatment of Huh7 cells profoundly enhances their differentiation state, thus improving the usefulness of this common cell line as an in vitro hepatocyte model.
Choi
S
S
Center for Pharmaceutical Biotechnology,University of Illinois at Chicago, 833 S. Wood Street, Chicago, IL 60612, USA.
Sainz
B
B
Corcoran
P
P
Uprichard
S
S
Jeong
H
H
D002454:Cell Differentiation / Q000187:drug effects*; D045744:Cell Line, Tumor; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism*; D017931:DNA Primers; D004121:Dimethyl Sulfoxide / Q000378:metabolism / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008748:Methylcholanthrene; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D008748:Methylcholanthrene; D019363:Cytochrome P-450 CYP1A1; D004121:Dimethyl Sulfoxide