title: Androgen responsiveness of Renilla luciferase reporter vectors is promoter, transgene, and cell line dependent.
abstract: 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.
authors.affiliation: The Prostate Centre, Jack Bell Research, Vancouver, BC, Canada. djm@interchange.ubc.ca
authors.forename: David J
authors.initials: DJ
authors.lastname: Mulholland
authors.forename: Michael
authors.initials: M
authors.lastname: Cox
authors.forename: Jason
authors.initials: J
authors.lastname: Read
authors.forename: Paul
authors.initials: P
authors.lastname: Rennie
authors.forename: Colleen
authors.initials: C
authors.lastname: Nelson
mesh_terms: D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D037421:Anthozoa / Q000201:enzymology / Q000235:genetics; D001681:Biological Assay; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D008297:Male; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000473:pathology; D015203:Reproducibility of Results; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D000728:Androgens; D008156:Luciferases