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MicroRNAs negatively regulate vitamin D receptor and cytochrome P450 3A4 enzyme pathway
Author(s) -
Pan Yuzhuo,
Gao WenQing,
Yu Aiming
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.757.2
Subject(s) - calcitriol receptor , microrna , luciferase , transfection , cyp3a4 , three prime untranslated region , untranslated region , microbiology and biotechnology , biology , cytochrome p450 , receptor , messenger rna , chemistry , enzyme , biochemistry , gene
CYP3A4 is an important hemoprotein that metabolizes many drugs. To understand posttranscriptional regulation of CYP3A4 and role of microRNAs (miRNAs) in drug metabolism, we found that two miRNAs might control posttranscriptional and transcriptional regulation of CYP3A4 through their actions on 3′UTR of CYP3A4 and vitamin D receptor (VDR/NR1I1). Luciferase assays showed that CYP3A4 3′UTR‐luciferase activity was decreased significantly after transfected with plasmid expressing miR‐27b or mmu‐miR‐298, whereas the activity was unchanged in cells transfected with miR‐122a or miR‐328. Disruption of corresponding miRNA response element (MRE) within CYP3A4 3′UTR led to 2‐ to 3‐fold increase in luciferase activity. Immunoblot analyses indicated that CYP3A4 protein expression was down‐regulated over 40% by miR‐27b and mmu‐miR‐298 in LS‐180 and PANC1 cells. Down‐regulation of VDR by the two miRNAs was supported by luciferase assay, immunoblot and qPCR analyses. Furthermore, Overexpression of miR‐27b or mmu‐miR‐298 resulted in lower cell sensitivity to cyclophosphamide. Together, these findings suggest that noncoding miRNAs may regulate CYP3A4 and VDR protein expression at the posttranscriptional level, and consequently affect drug‐metabolizing capacity. This project was supported by UB Interdisciplinary Research Development Fund, and in part by NIH R01DA021172.