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The Modified Firefly Luciferase Reporter Gene (luc+) but not Renilla Luciferase is Induced by all-trans Retinoic Acid in MCF-7 Breast Cancer Cells

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Abstract

Luciferase genes are widely used as reporters to analyze promoter and regulatory elements. We found that a luciferase reporter gene vector with a modified firefly luciferase gene (luc+), but not Renilla luciferase (Rluc), was induced by all-trans retinoic acid (tRA) in the MCF-7 breast cancer cell line. tRA (5 × 10−6 M) increased luciferase activity of the pGL3 promoter vector (containing luc+) up to ∼3.8-fold in MCF-7 cells, but not in LNCaP prostate cancer cells or JEG-3 choriocarcinoma cells. Chimeric plasmids were constructed and showed that tRA-induction required the luc+ gene, but not any specific promoter or vector sequence. Time course and dose-response studies of tRA-induction indicated that longer treatment (>24 h) and higher tRA dose (>10−6 M) were required for luc+ induction compared with those for a positive retinoic acid response element (maximum induction at 6 h and 10−8 M tRA). Studies with the translation inhibitor, cycloheximide, indicated the half-life of the luc+ protein was increased from 9.7 ± 1.5 to 22.1 ± 3.1 h with tRA treatment. Other retinoids, TTNPB, a retinoic acid receptor β/γ-specific ligand, and a retinoid X receptor ligand, did not significantly increase luc+ expression. Caution is needed in analysis of retinoid responsive gene regulation with the luciferase reporter system in MCF-7 cells, especially at high retinoid concentrations.

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Authors and Affiliations

  1. Molecular Endocrinology Laboratory, VA Greater Los Angeles Healthcare System, Departments of Medicine and Physiology, UCLA School of Medicine, Los Angeles, CA, USA

    Takahiko Kogai, Yoko Kanamoto & Gregory A. Brent

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  1. Takahiko Kogai
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  2. Yoko Kanamoto
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  3. Gregory A. Brent
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Kogai, T., Kanamoto, Y. & Brent, G.A. The Modified Firefly Luciferase Reporter Gene (luc+) but not Renilla Luciferase is Induced by all-trans Retinoic Acid in MCF-7 Breast Cancer Cells. Breast Cancer Res Treat 78, 119–126 (2003). https://doi.org/10.1023/A:1022179717847

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  • DOI: https://doi.org/10.1023/A:1022179717847

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