A nuclear binding assay for measurement of biologically active androgen receptors in animal tissues and human prostate cancer.
A nuclear binding (NB) assay has been developed for the measurement in intact viable cells of biologically active (functional) estrogen and progesterone receptors, i.e. those capable of binding to nuclear acceptor sites [Spelsberg et al., Endocrinology 121: 631 (1987)]. This paper describes the application of this assay to analyses of androgen receptors in the guinea pig seminal vesicle and in human prostatic carcinoma. Cells from fresh animal seminal vesicles or human prostate carcinoma are isolated using collagenase and are incubated with [3H]R1881 for 1 h at 22 degrees C, after which nuclei are isolated at 4 degrees C and assayed for DNA and radioactivity. This NB assay demonstrates a saturable, temperature dependent, steroid and tissue specific nuclear binding of [3H]R1881 for the guinea pig-seminal vesicle system. The nuclear binding is of high affinity and low capacity. The NB assay reveals several important aspects of the androgen and estrogen receptors in target tissues: (1) the nuclear acceptor sites for androgen receptor (AR) are steroid receptor specific; (2) there are different concentrations of the androgen and estrogen receptors between the epithelium and the fibromuscular components of the guinea pig seminal vesicle; and finally (3) some biopsies of human prostate cancer appear to contain biologically inactive AR. This assay may be useful in the analyses of functional receptors in biopsies of human cancer cells.
Umehara
T
T
Department of Biochemistry and Molecular Biology, Mayo Graduate School of Medicine, Mayo Clinic, Rochester, MN 55905.
Graham
M L
ML
Berg
N J
NJ
Lieber
M M
MM
Spelsberg
T C
TC
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D004963:Estrenes / Q000378:metabolism; D006168:Guinea Pigs; D006801:Humans; D008297:Male; D015741:Metribolone; D011471:Prostatic Neoplasms / Q000378:metabolism*; D011944:Receptors, Androgen / Q000378:metabolism*; D012669:Seminal Vesicles / Q000378:metabolism
D004963:Estrenes; D011944:Receptors, Androgen; D015741:Metribolone