Hormone-dependent androgen receptor phosphorylation is accompanied by receptor transformation in human lymph node carcinoma of the prostate cells

Phosphorylation of the androgen receptor was investigated in the absence of hormone as well as during and after transformation of the receptor to the tight nuclear binding form. Human prostate tumor cells (LNCaP) were labeled for 4 h with [32P]orthophosphate in the presence or absence of steroid. Su...

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Bibliographic Details
Published in:The Journal of biological chemistry 1991-02, Vol.266 (6), p.3734-3738
Main Authors: van Laar, J H, Berrevoets, C A, Trapman, J, Zegers, N D, Brinkmann, A O
Format: Article
Language:English
Subjects:
Biological and medical sciences
carcinoma
Cell receptors
Cell structures and functions
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors
Hormones - metabolism
Humans
Ligands
lymph nodes
Lymphatic Metastasis
Male
Molecular and cellular biology
Phosphorylation
Precipitin Tests
prostate
Prostatic Neoplasms - metabolism
Prostatic Neoplasms - pathology
Receptors, Androgen - metabolism
Substrate Specificity
Tumor Cells, Cultured
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Summary:Phosphorylation of the androgen receptor was investigated in the absence of hormone as well as during and after transformation of the receptor to the tight nuclear binding form. Human prostate tumor cells (LNCaP) were labeled for 4 h with [32P]orthophosphate in the presence or absence of steroid. Subsequently, androgen receptors were immunoprecipitated either from total cell lysates or from nuclear extracts using a specific monoclonal antibody. The immunoprecipitated receptor preparations were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blotting, using a polyclonal antiserum, and autoradiography. It was observed that the androgen receptor is already phosphorylated in the absence of hormone, but undergoes a hormone-induced additional phosphorylation. After administration of 10 nM R1881, a 1.8-fold increase in phosphorylation over nonstimulated control cells was reached. Moreover, the amount of nuclear extractable androgen receptor was increased; the acquisition of tight nuclear binding capacity was accompanied by hormone-induced receptor phosphorylation.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)67856-7