Induction of UDP-glucuronosyltransferase 1A8 mRNA by 3-methylcholanthene in rat hepatoma cells.

UDP-glucuronosyltransferases (UGTs) catalyze the glucuronidation of a broad spectrum of endobiotic and xenobiotic compounds, which leads to the excretion of hydrophilic glucuronides via bile or urine. By a mechanism of exon sharing, isoforms of the UGT1 family are made from the complex gene locus by an alternative combination of one of the unique first exons with the other commonly used exons. This study demonstrates that the expression of the UGT1 gene UGT1A6, 1A7 and 1A8 is regulated at the transcriptional level by 3-methylcholanthene (3-MC) in rat hepatoma H-4-II-E cells. Following 3-MC treatment, there is a gradual increase in the amount of UGT1A6 and UGT1A7 mRNA to the maximum levels after 16hr of treatment. The induction effect of 3-MC led to the expression of UGT1A8 which has not been reported before. This induction is suppressed by the RNA synthesis inhibitor actinomycin D, indicating that the inducer does not act at the level of mRNA stabilization. Northern blot analysis showed a 4-fold increase in UGT1A8 transcription after treatment with 3-MC. The prolonged treatment with the protein synthesis inhibitor did not affect the induction process. The results provide experimental evidence for a transcriptional control of UGT1A8 synthesis. Transcriptional activation of the UGT1A8 by 3-MC does not appear to require de novo protein synthesis. 3-MC dependent activation is probably the result of a direct action of the compound on the aryl hydrocarbon receptor complex (AhR).