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Androgen responsiveness of Renilla luciferase reporter vectors is promoter, transgene, and cell line dependent
Author(s) -
Mulholland David J.,
Cox Michael,
Read Jason,
Rennie Paul,
Nelson Colleen
Publication year - 2004
Publication title -
the prostate
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.295
H-Index - 123
eISSN - 1097-0045
pISSN - 0270-4137
DOI - 10.1002/pros.20059
Subject(s) - luciferase , lncap , androgen receptor , biology , dihydrotestosterone , transfection , androgen , transgene , reporter gene , microbiology and biotechnology , cell culture , glucocorticoid receptor , receptor , prostate cancer , gene expression , endocrinology , gene , biochemistry , genetics , hormone , cancer
Renilla based reporters are frequently used as transfection controls for luciferase transcriptional reporter assays. However, recent evidence suggests that a commonly used reporter (HSV‐thymidine kinase driven Renilla ) is responsive to androgen receptor (AR) and glucocorticoid receptors in the presence of the cognate ligands, dihydrotestosterone (DHT) and dexamethasone (DEX), respectively [1]. We further validate this important technical difficulty by illustrating that in LNCaP prostate cancer cells, spurious Renilla luciferase activity is a function of (a) the promoter driving Renilla expression, (b) the presence of co‐transfected transgenes, and (c) the androgen responsiveness of the cell line used. Using inhibitors of transcription and translation we showed that transcript interference or translational modulation is not a major means by which androgens affect Renilla luciferase activity. As luciferase reporter assays are a frequent means of studying transcriptional co‐regulation in the highly androgen dependent LNCaP cell line, our data serves as a cautionary note that alternative normalization techniques should be employed to avoid misinterpretation of data. © 2004 Wiley‐Liss, Inc.