pp 1-9 | Cite as

Detection of Total UDP-Glucuronosyltransferase (UGT) Activity in Melanoma Cells

  • Ryan W. Dellinger
  • Frank L. MeyskensJr.
Part of the Methods in Molecular Biology book series


The UDP-glucuronosyltransferases (UGTs) are integrally involved in the clearance of a wide range of drugs used to combat human diseases. UGT expression levels and activity can be induced by drug addition to cells and has been proposed as a potential intratumoral drug resistance mechanism. Traditional methods of assaying UGT activity are drug-centric and require HPLCs with multiple detectors (dependent on individual drug). Here, we describe a generalized method to detect total UGT activity (intrinsic or induced) via the UGT-Glo assay which utilizes a general UGT substrate with luminescence as the readout eliminating the need for multiple HPLC detectors to detect total UGT activity in a given sample. The method detailed here can be applied for any UGT containing sample, allowing for the efficient detection of total UGT activity to be the functional endpoint using a plate reader. In this manner, global changes in UGT activity can be monitored in response to a wide variety of stimuli.


UDP-glucuronosyltransferase Glucuronidase activity UGT-Glo Melanoma Drug resistance 


  1. 1.
    Nagar S, Remmel RP (2006) Uridine diphosphoglucuronosyltransferase pharmacogenetics and cancer. Oncogene 25:1659–1672PubMedCrossRefGoogle Scholar
  2. 2.
    Tukey RH, Strassburg CP (2000) Human UDP-glucuronosyltransferases: metabolism, expression, and disease. Annu Rev Pharmacol Toxicol 40:581–616PubMedCrossRefGoogle Scholar
  3. 3.
    Chouinard S, Yueh MF, Tukey RH et al (2008) Inactivation by UDP-glucuronosyltransferase enzymes: the end of androgen signaling. J Steroid Biochem Mol Biol 109:247–253PubMedCrossRefGoogle Scholar
  4. 4.
    Starlard-Davenport A, Lyn-Cook B, Radominska-Pandya A (2008) Identification of UDP-glucuronosyltransferase 1A10 in non-malignant and malignant human breast tissues. Steroids 73:611–620PubMedPubMedCentralCrossRefGoogle Scholar
  5. 5.
    Gregory PA, Lewinsky RH, Gardner-Stephen DA, Mackenzie PI (2004) Regulation of UDP glucuronosyltransferases in the gastrointestinal tract. Toxicol Appl Pharmacol 199:354–363PubMedCrossRefGoogle Scholar
  6. 6.
    Giuliani L, Ciotti M, Stoppacciaro A et al (2005) UDP-glucuronosyltransferases 1A expression in human urinary bladder and colon cancer by immunohistochemistry. Oncol Rep 13:185–191PubMedGoogle Scholar
  7. 7.
    King CD, Rios GR, Green MD, Tephly TR (2000) UDP-glucuronosyltransferases. Curr Drug Metab 1:143–161PubMedCrossRefGoogle Scholar
  8. 8.
    Zheng Z, Fang JL, Lazarus P (2002) Glucuronidation: an important mechanism for detoxification of benzo[a]pyrene metabolites in aerodigestive tract tissues. Drug Metab Dispos 30:397–403PubMedCrossRefGoogle Scholar
  9. 9.
    Dellinger RW, Fang JL, Chen G, Weinberg R, Lazarus P (2006) Importance of UDP-glucuronosyltransferase 1A10 (UGT1A10) in the detoxification of polycyclic aromatic hydrocarbons: decreased glucuronidative activity of the UGT1A10139Lys isoform. Drug Metab Dispos 34:943–949PubMedGoogle Scholar
  10. 10.
    Dellinger RW, Matundan HH, Ahmed AS, Duong PH, Meyskens FL Jr (2012) Anti-cancer drugs elicit re-expression of UDP-glucuronosyltransferases in melanoma cells. PLoS One 7, e47696PubMedADSPubMedCentralCrossRefGoogle Scholar
  11. 11.
    Bansal SK, Gessner T (1980) A unified method for the assay of uridine diphosphoglucuronyltransferase activities toward various aglycones using uridine diphospho[U-14C]glucuronic acid. Anal Biochem 109:321–329PubMedCrossRefGoogle Scholar
  12. 12.
    Fisher MB, Campanale K, Ackermann BL, VandenBranden M, Wrighton SA (2000) In vitro glucuronidation using human liver microsomes and the pore-forming peptide alamethicin. Drug Metab Dispos 28:560–566PubMedGoogle Scholar
  13. 13.
    Dellinger RW, Chen G, Blevins-Primeau AS, Krzeminski J, Amin S, Lazarus P (2007) Glucuronidation of PhIP and N-OH-PhIP by UDP-glucuronosyltransferase 1A10. Carcinogenesis 28:2412–2418PubMedCrossRefGoogle Scholar
  14. 14.
    Radominska-Pandya A, Pokrovskaya ID, Xu J et al (2002) Nuclear UDP-glucuronosyltransferases: identification of UGT2B7 and UGT1A6 in human liver nuclear membranes. Arch Biochem Biophys 399:37–48PubMedCrossRefGoogle Scholar
  15. 15.
    Chen G, Dellinger RW, Gallagher CJ, Sun D, Lazarus P (2008) Identification of a prevalent functional missense polymorphism in the UGT2B10 gene and its association with UGT2B10 inactivation against tobacco-specific nitrosamines. Pharmacogenet Genomics 18:181–191PubMedCrossRefGoogle Scholar
  16. 16.
    Dellinger RW, Garcia AM, Meyskens FL Jr (2014) Differences in the glucuronidation of resveratrol and pterostilbene; altered enzyme specificity and potential gender differences. Drug Metab Pharmacokinet 29(2):112–119PubMedPubMedCentralCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Chao Family Comprehensive Cancer Center and Departments of Medicine (RWD, FLM), Biological Chemistry (FLM), Epidemiology (RWD) and Public Health (FLM)University of California IrvineIrvineUSA

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