A mini-column method for routine measurement of human prostatic androgen receptors.
The complex heterogeneous nature of the human prostate gland is such that it is advisable to know the histological characteristics of each sample used for androgen receptor (AR) measurement. Adequate size of sample for AR determination is thus a problem if specimens provided during routine transurethral prostatectomy are to be used for both estimation of AR and histological examination. We present a simple method suitable for these small specimens in which [3H]R 1881 bound to AR is separated from free steroid on mini-columns of controlled-pore glass beads. Data obtained indicate a single class of binding sites of high affinity and low capacity with steroid specificity typical of an androgen receptor. The assay is suitable for samples as small as 20 mg wet weight and is linear using 25-125 microliter cytosol (correlation coefficient 0.995). Intra-assay variation is 6.8% and interassay variation 25.8% (n = 22) over 4 months. A single saturating concentration of steroid measures 97% of AR calculated by Scatchard analysis. Inclusion of high salt (0.4 M KNO3) and 10 mM dithiothreitol in incubation buffer at pH 8.4 are essential; inclusion of 10 mM sodium molybdate in the homogenisation buffer improves measurement. A comparison of AR measured in histologically similar samples obtained by a transurethral resectoscope (TUR) and a cold punch resectoscope (CPR) taken in juxtaposition demonstrated no difference in receptor content. Although carcinomatous samples contained significantly higher receptors levels than benign samples, no differences were observed between TUR and CPR specimens.
D002850:Chromatography, Gel; D003600:Cytosol / Q000032:analysis; D004229:Dithiothreitol / Q000494:pharmacology; D004963:Estrenes / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D008297:Male; D015741:Metribolone; D008982:Molybdenum / Q000494:pharmacology; D011467:Prostate / Q000032:analysis*; D011944:Receptors, Androgen / Q000032:analysis*; D013048:Specimen Handling
D004963:Estrenes; D011944:Receptors, Androgen; C044659:molybdate; D015741:Metribolone; D008982:Molybdenum; D004229:Dithiothreitol