Potential Regulation of UGT2B10 and UGT2B7 by miR-485-5p in Human Liver. Other Scholarly Work

Sutliff, Aimee K, Shi, Jian, Watson, Christy JW et al. (2019). Potential Regulation of UGT2B10 and UGT2B7 by miR-485-5p in Human Liver. . 96(6), 674-682. 10.1124/mol.119.115881

cited authors

  • Sutliff, Aimee K; Shi, Jian; Watson, Christy JW; Hunt, Martina S; Chen, Gang; Zhu, Hao-Jie; Lazarus, Philip

fiu authors

abstract

  • The UDP-glucuronosyltransferase (UGT) family of enzymes is important in the metabolic elimination of a variety of endogenous compounds such as bile acids, steroids, and fat-soluble vitamins, as well as exogenous compounds including many pharmaceuticals. The UGT2B subfamily is a major family of UGT enzymes expressed in human liver. The identification of novel mechanisms including post-transcriptional regulation by microRNA (miRNA) contributes to interindividual variability in UGT2B expression and is a crucial component in predicting patient drug response. In the present study, a high-resolution liquid chromatography-tandem mass spectrometry method was employed to measure UGT2B protein levels in a panel of human liver microsomal samples (n = 62). Concurrent in silico analysis identified eight candidate miRNAs as potential regulators of UGT2B enzymes. Comparison of UGT2B protein expression and candidate miRNA levels from human liver samples demonstrated a significant inverse correlation between UGT2B10 and UGT2B15 and one of these candidate miRNAs, miR-485-5p. A near-significant correlation was also observed between UGT2B7 and miR-485-5p expression. In vitro analysis using luciferase-containing vectors suggested an interaction of miR-485-5p within the UGT2B10 3'-untranslated region (UTR), and significant reduction in luciferase activity was also observed for a luciferase vector containing the UGT2B7 3'-UTR; however, none was observed for the UBT2B15 3'-UTR. UGT2B10 and UGT2B7 activities were probed using nicotine and 3'-azido-3'-deoxythymidine, respectively, and significant decreases in glucuronidation activity were observed for both substrates in HuH-7 and Hep3B cells upon overexpression of miR-485-5p mimic. This is the first study demonstrating a regulatory role of miR-485-5p for multiple UGT2B enzymes. SIGNIFICANCE STATEMENT: The purpose of this study was to identify novel epigenetic miRNA regulators of the UGT2B drug-metabolizing enzymes in healthy human liver samples. Our results indicate that miRNA 485-5p is a novel regulator of UGT2B7 and UGT2B10, which play an important role in the metabolism of many commonly prescribed medications, carcinogens, and endogenous compounds. This study identified potential miRNA-UGT2B mRNA interactions using a novel proteomic approach, with in vitro experiments undertaken to validate these interactions.

publication date

  • December 1, 2019

keywords

  • Glucuronosyltransferase
  • HEK293 Cells
  • Humans
  • Liver
  • MicroRNAs
  • Proteomics

Digital Object Identifier (DOI)

Medium

  • Print-Electronic

start page

  • 674

end page

  • 682

volume

  • 96

issue

  • 6