Regulation of UGT2B Expression and Activity by miR-216b-5p in Liver Cancer Cell Lines. Other Scholarly Work

Dluzen, Douglas F, Sutliff, Aimee K, Chen, Gang et al. (2016). Regulation of UGT2B Expression and Activity by miR-216b-5p in Liver Cancer Cell Lines. . 359(1), 182-193. 10.1124/jpet.116.235044

cited authors

  • Dluzen, Douglas F; Sutliff, Aimee K; Chen, Gang; Watson, Christy JW; Ishmael, Faoud T; Lazarus, Philip

fiu authors

abstract

  • The UDP-glucuronosyltransferase (UGT) 2B enzymes are important in the detoxification of a variety of endogenous and exogenous compounds, including many hormones, drugs, and carcinogens. Identifying novel mechanisms governing their expression is important in understanding patient-specific response to drugs and cancer risk factors. In silico prediction algorithm programs were used to screen for microRNAs (miRNAs) as potential regulators of UGT2B enzymes, with miR-216b-5p identified as a potential candidate. Luciferase data suggested the presence of a functional miR-216b-5p binding motif within the 3' untranslated regions of UGTs 2B7, 2B4, and 2B10. Overexpression of miR-216b-5p mimics significantly repressed UGT2B7 (P < 0.001) and UGT2B10 (P = 0.0018) mRNA levels in HuH-7 cells and UGT2B4 (P < 0.001) and UGT2B10 (P = 0.018) mRNA in Hep3B cells. UGT2B7 protein levels were repressed in both HuH-7 and Hep3B cells in the presence of increasing miR-216b-5p concentrations, corresponding with significant (P < 0.001 and P = 0.011, respectively) decreases in glucuronidation activity against the UGT2B7-specific substrate epirubicin. Inhibition of endogenous miR-216b-5p levels significantly increased UGT2B7 mRNA levels in HuH-7 (P = 0.021) and Hep3B (P = 0.0068) cells, and increased epirubicin glucuronidation by 85% (P = 0.057) and 50% (P = 0.012) for HuH-7 and Hep3B cells, respectively. UGT2B4 activity against codeine and UGT2B10 activity against nicotine were significantly decreased in both HuH-7 and Hep3B cells (P < 0.001 and P = 0.0048, and P = 0.017 and P = 0.043, respectively) after overexpression of miR-216b-5p mimic. This is the first evidence that miRNAs regulate UGT 2B7, 2B4, and 2B10 expression, and that miR-216b-5p regulation of UGT2B proteins may be important in regulating the metabolism of UGT2B substrates.

publication date

  • October 1, 2016

keywords

  • 3' Untranslated Regions
  • Base Sequence
  • Cell Line, Tumor
  • Gene Expression Regulation, Neoplastic
  • Glucuronosyltransferase
  • Humans
  • Liver Neoplasms
  • MicroRNAs
  • Polymorphism, Genetic

Digital Object Identifier (DOI)

Medium

  • Print-Electronic

start page

  • 182

end page

  • 193

volume

  • 359

issue

  • 1