Abstract
It is widely believed that the UGT1A isoforms, UGT1A8 and -1A10, are expressed exclusively in extrahepatic tissues. In this work, human primary hepatocytes from six donors were analyzed for UGT1A8 and -1A10 mRNA expression by semi-quantitative RT-PCR. New primers to amplify UGT1A8 mRNA were designed and found to differ from those previously published. We demonstrated that UGT1A8 and -1A10 mRNA are expressed in hepatocytes. Although basal UGT mRNA levels were detected in untreated hepatocytes, significant up-regulation of the levels of mRNA for these isoforms were seen after treatment with 3-methylcholanthrene (3-MC) and rifampicin (Rif). RT-PCR products for all UGTs were sequenced and unambiguously identified as matching the corresponding cDNA. The discovery of these isoforms in hepatocytes is a novel discovery and will stimulate studies on the potential role for these isoforms in hepatic detoxification.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Adult
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Aged
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Base Sequence
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Cells, Cultured
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Child, Preschool
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Female
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Gastrointestinal Tract / metabolism
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Gene Expression Regulation, Enzymologic / drug effects
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Glucuronosyltransferase / genetics*
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Hepatocytes / cytology
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Hepatocytes / drug effects
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Hepatocytes / metabolism*
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Humans
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Intestinal Mucosa / metabolism
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Isoenzymes / genetics
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Male
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Methylcholanthrene / pharmacology
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Middle Aged
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Molecular Sequence Data
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Polymerase Chain Reaction
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RNA, Messenger / genetics
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RNA, Messenger / metabolism*
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Reverse Transcriptase Polymerase Chain Reaction
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Rifampin / pharmacology
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Sequence Homology, Nucleic Acid
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UDP-Glucuronosyltransferase 1A9
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Up-Regulation / drug effects
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Up-Regulation / genetics
Substances
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Isoenzymes
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RNA, Messenger
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Methylcholanthrene
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bilirubin uridine-diphosphoglucuronosyl transferase 1A10
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Glucuronosyltransferase
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UDP-Glucuronosyltransferase 1A9
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UDP-glucuronosyltransferase, UGT1A8
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Rifampin