Relationship between mRNA Levels Quantified by Reverse Transcription-Competitive PCR and Metabolic Activity of CYP3A4 and CYP2E1 in Human Liver

Reverse transcription-competitive polymerase chain reaction is a powerful and sensitive tool for quantifying the absolute amount of mRNA. Using this method with β-actin as the standard, we measured the mRNA level of CYP3A and CYP2E1 isoforms in human livers. We also determined the metabolic activiti...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 1999-08, Vol.262 (2), p.499-503
Main Authors: Sumida, Akihiko, Kinoshita, Kayoko, Fukuda, Tsuyoshi, Matsuda, Hideyasu, Yamamoto, Isamu, Inaba, Tadanobu, Azuma, Junichi
Format: Article
Language:English
Subjects:
Adolescent
Adult
CYP2E1
CYP3A4
Cytochrome P-450 CYP2E1 - analysis
Cytochrome P-450 CYP2E1 - genetics
Cytochrome P-450 CYP3A
Cytochrome P-450 Enzyme System - analysis
Cytochrome P-450 Enzyme System - genetics
European Continental Ancestry Group
Female
human liver
Humans
Male
metabolic activity
Microsomes, Liver - enzymology
Middle Aged
Mixed Function Oxygenases - analysis
Mixed Function Oxygenases - genetics
mRNA
Reverse Transcriptase Polymerase Chain Reaction
reverse transcription-competitive PCR
RNA, Messenger - analysis
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Summary:Reverse transcription-competitive polymerase chain reaction is a powerful and sensitive tool for quantifying the absolute amount of mRNA. Using this method with β-actin as the standard, we measured the mRNA level of CYP3A and CYP2E1 isoforms in human livers. We also determined the metabolic activities for both CYP isoforms. The absolute amounts of CYP3A4 and CYP2E1 mRNA in 15 liver tissues ranged from 15 to 5127 and 15163 to 69289 copies/104 copies of β-actin (341- & 3.6-fold), respectively. The testosterone 6β-hydroxylation for CYP3A4 and chlorzoxazone 6-hydroxylation activity for CYP2E1 ranged from 30 to 505 pmol/mg/min (16-fold) and from 0.59 to 2.73 nmol/mg/ml (3.6-fold), respectively. The correlation between the mRNA level and activity of CYP3A4 was significant (r = 0.94), while there was no significant correlation for CYP2E1 (r = 0.04). In conclusion, we observed a significant correlation between enzyme activity and mRNA expression for CYP3A4 but not for CYP2E1. This fact indicates that CYP2E1, in addition to being less variable between individuals than CYP3A4, differs in its regulation mechanism.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1999.1233