Modeling of Androgen Receptor Activated Gene Transcription in Nervous System
Date Issued
2005
Date
2005
Author(s)
Fu, Yin-Chieh
DOI
en-US
Abstract
Androgens are steroids critical for the development and maintenance of the male phenotype. Androgens are known to regulate functions of various tissues, including muscular tissue, nervous system, bone, and sexual behavior. Testosterone (T) and its metabolite dihydrotestosterone (DHT) are well-known androgens exert their effects through binding to the androgen receptor (AR), which is a transcription factor. Hormone binding promotes the nuclear translocation of the AR, where AR recognizes certain DNA sequence called androgen response element (ARE) and regulates gene expression. AR positively regulates the transcription of prostate specific antigen (PSA) mainly through binding to AREs in the proximal region and the PSA distal enhancer region of PSA promoter. To understand androgen functions in the nervous system, we introduced GFP, reporter gene, regulated by PSA promoter into zebrafish embryos. Examine GFP expression in zebrafish embryos by using fluorescent microscope, revealing that PSA1.5 promoter expresses significantly in notochord, muscle, and neuronal tissues. Androgens not only play an important role in reproductive tissues, but also in nervous system during neurogenesis. Indeed, neuronal expression of PSA promoter is further confirmed by using transient transfection in different human neuronal cell lines. We want to set up a model to study AR mediated genomic effect in nervous cells by using PSA1.5 promoter as a tool. Here we choose human neuroblastoma cell line SK-N-BE(2) as an in vitro model because it may differentiate into neuron-like phenotype and has been studied in estrogen receptor regulated neuron differentiation. Human AR are introduced into SK-N-BE(2) cells by stable transfection derived SK-AR62 line. Further characterization showed that SK-AR62 grows in an androgen-dependent manner. However, anti-androgen cannot inhibit DHT induced cell proliferation in SK-AR62. Stable transfection of PSA1.5-Luc into SK-AR62 results in constitutive expression of luciferase disregard of hormone treatment. Furthermore, introduction of plasmids harboring AR expression cassette and PSA1.5-Luc reporter into SK-N-BE(2) to derive SK-PLAR clones, results in constitutive expression of luciferase as aforementioned result. Among the SK-PLAR clones studied, SK-PLAR54 can differentiate neuron-like morphology upon 3 days culture in serum free medium. Overall, these results indicate PSA1.5-Luc expresses androgen-independently in the presence of AR in SK-N-BE(2) derived cells. The AR may activate the 1.5Kb PSA promoter ligand-independently in SK-N-BE(2) cells.
Subjects
雄性素受體
前列腺特異抗原
神經系統
androgen receptor
PSA
SK-N-BE(2)
nervous system
Type
other
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