The modified firefly luciferase reporter gene (luc+) but not Renilla luciferase is induced by all-trans retinoic acid in MCF-7 breast cancer cells

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 l...

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Bibliographic Details
Published in:Breast cancer research and treatment 2003-03, Vol.78 (1), p.119-126
Main Authors: KOGAI, Takahiko, KANAMOTO, Yoko, BRENT, Gregory A
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Breast cancer
Breast Neoplasms - genetics
Cancer research
Cancer therapies
Coleoptera
Dose-Response Relationship, Drug
Female
Gene Expression Regulation, Neoplastic - drug effects
Genes, Reporter - drug effects
Genes, Reporter - genetics
Genetic Vectors
Gynecology. Andrology. Obstetrics
Humans
Luciferases - drug effects
Luciferases - genetics
Mammary gland diseases
Medical sciences
Tretinoin - pharmacology
Tretinoin - physiology
Tumor Cells, Cultured
Tumors
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Summary: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 x 10(-6) M) increased luciferase activity of the pGL3 promoter vector (containing luc+) up to approximately 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 (> 24h) 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 beta/gamma-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.
ISSN:0167-6806
1573-7217
DOI:10.1023/A:1022179717847