Superinduction of estrogen receptor mediated gene expression in luciferase based reporter gene assays is mediated by a post-transcriptional mechanism

A.M. Sotoca Covaleda, T.F.H. Bovee, W. Brand, N.R. Velikova, A.J. Murk, J.J.M. Vervoort, I.M.C.M. Rietjens

Research output: Contribution to journalArticleAcademicpeer-review

40 Citations (Scopus)

Abstract

Several estrogenic compounds including the isoflavonoid genistein have been reported to induce a higher maximal response than the natural estrogen 17ß-estradiol in in vitro luciferase based reporter gene bioassays for testing estrogenicity. The phenomenon has been referred to as superinduction. The mechanism underlying this effect and thus also its biological relevance remain to be elucidated. In the present study several hypotheses for the possible mechanisms underlying this superinduction were investigated using genistein as the model compound. These hypotheses included (i) a non-estrogen receptor (ER)-mediated mechanism, (ii) a role for an ER activating genistein metabolite with higher ER inducing activity than genistein itself, and (iii) a post-transcriptional mechanism that is not biologically relevant but specific for the luciferase based reporter gene assays. The data presented in this study indicate that induction and also superinduction of the reporter gene is ER-mediated, and that superinduction by genistein could be ascribed to stabilization of the firefly luciferase reporter enzyme increasing the bioluminescent signal during the cell-based assay. This indicates that the phenomenon of superinduction may not be biologically relevant but may rather represent a post-transcriptional effect on enzyme stability.
Original languageEnglish
Pages (from-to)204-211
JournalJournal of Steroid Biochemistry and Molecular Biology
Volume122
Issue number4
DOIs
Publication statusPublished - 2010

Keywords

  • human breast-cancer
  • in-vitro
  • er-beta
  • firefly luciferase
  • cell-proliferation
  • genistein
  • phytoestrogens
  • metabolites
  • daidzein
  • alpha

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