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Role of Multiple MicroRNAs in the Sexually Dimorphic Expression.ppt
Special Sectionon Epigenetic Regulation of Drug MetabolizingEnzymes and Transporters Role of Multiple MicroRNAs in the Sexually Dimorphic Expression of Cyp2b9 in Mouse Liver ABSTRACT In this study, we found evidence that some microRNAs contributed to the sexually biased expression of Cyp2b9 via post-transcriptional regulation. Luciferase assays revealed that approximate half of these microRNAs repressed luciferase activity in a reporter system containing the 3’-untranslated region (3’-UTR) of Cyp2b9, and also inhibited Cyp2b9 protein expression. MicroRNA seed region mutation or mutations in putative binding sites of the microRNAs in Cyp2b9 3’-UTR led to the loss of the suppression of luciferase activity. There was also a negative correlation between the levels of these microRNAs and Cyp2b9. Our results suggested that multiple microRNAs participated in the regulation of Cyp2b9 expression, and that the lower expression levels of these microRNAs potentially contributed to the female-specific expression of Cyp2b9 in the livers of C57BL/6J mice. Introduction The cytochrome P450 (P450) superfamily is a large and diverse group of enzymes that catalyze the metabolism of drugs and carcinogens. Cyp2b9 is a testosterone 16a-hydroxylase enzyme showing female-specific expression in many inbred mouse strains, including C57BL/6J (Noshiro et al., 1988). Previous studies have identified some mechanisms involved in the sexually dimorphic expression of Cyp2b9 at the transcriptional level. MicroRNAs are a large family of endogenous noncoding RNAs that regulate gene expression primarily by binding to the 3’-untranslatedregion (3’-UTR) of target genes, resulting in suppressed translation or decreased mRNA stability. Introduction Dicer1 is an RNase III endonuclease that is essential for the biogenesis of microRNAs, and multiple Dicer1knockout (KO) animal models show significant down-regulation of microRNAs and upregulation of microRNA targeting genes in these models (Hand et al.,
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