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Biochem Pharmacol. 2017 Sep 15;140:139-149. doi: 10.1016/j.bcp.2017.05.018. Epub 2017 May 26.

MicroRNA hsa-miR-370-3p suppresses the expression and induction of CYP2D6 by facilitating mRNA degradation.

Author information

1
Department of Oncology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai 519000, China; National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA.
2
National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA; Department of Gastroenterology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, Guangzhou 510120, China; Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China.
3
National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA.
4
School of Life Sciences, East China Normal University, Shanghai 200241, China.
5
Longevity Center, CHI St. Vincent Hospital, Little Rock, AR 72205, USA.
6
Department of Gastroenterology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, Guangzhou 510120, China; Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China.
7
National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA; School of Public Health, Qingdao University, Qingdao 266071, China. Electronic address: dianke.yu@fda.hhs.gov.
8
National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA. Electronic address: baitang.ning@fda.hhs.gov.

Abstract

Cytochrome P450 2D6 (CYP2D6) participates in the metabolism of approximately 20-25% of prescribed drugs. Genetic polymorphisms influence the expression and/or activity of CYP2D6, and inter-individual differences in drug activation and elimination caused by CYP2D6 genetic variants were reported. However, little is known about the potential modulation of CYP2D6 expression by microRNAs (miRNAs). In the current study, by using in silico prediction of the stabilities of miRNA/mRNA complexes, we screened 38 miRNA candidates that may interact with the transcript of CYP2D6. An inverse correlation between the expression of miRNA hsa-miR-370-3p and the expression of CYP2D6 was observed in human liver tissue samples. Electrophoretic mobility shift assays confirmed that hsa-miR-370-3p was able to directly bind to its cognate target within the coding region of the CYP2D6 transcript. The transfection of hsa-miR-370-3p mimics into the HepG2CYP2D6 cell line, a genetically modified cell line that overexpresses exogenous CYP2D6, was able to suppress the expression of CYP2D6 significantly at both mRNA and protein levels. The transfection of hsa-miR-370-3p mimics was also able to inhibit endogenous mRNA expression and/or protein production of CYP2D6 in HepaRG cells. Furthermore, in HepaRG, HepG2, and Huh7 cells, dexamethasone-induced expression of CYP2D6 was inhibited by hsa-miR-370-3p mimics. To investigate whether the miRNA mediated suppression is caused by inhibiting protein translation or promoting mRNA degradation, an actinomycin D assay was used to measure the stability of CYP2D6 transcripts. The results indicated that hsa-miR-370-3p mimics facilitated significantly the degradation of CYP2D6 mRNA. In addition, proteomics analyses of proteins isolated from the miRNA/mRNA/protein complex suggested that a group of multifunctional proteins facilitated the interaction between hsa-miR-370-3p and CYP2D6, thereby promoting mRNA degradation.

KEYWORDS:

CYP2D6; Drug metabolizing enzymes; Epigenetics; Pharmacogenomics; hsa-miR-370-3p

PMID:
28552654
PMCID:
PMC5687086
DOI:
10.1016/j.bcp.2017.05.018
[Indexed for MEDLINE]
Free PMC Article

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