Berns E M, de Boer W, Mulder E
Prostate. 1986;9(3):247-59. doi: 10.1002/pros.2990090305.
Hormone sensitivity, as indicated by the presence of steroid hormone receptors and by the effect of androgens and anti-androgens on the release of proteins by cultured cells of the human prostate tumor cell line lymph node carcinoma of the prostate-fast growing colony (LNCaP-FGC), has been studied. The growth of the LNCaP-FGC cells were stimulated by androgens in a dose-dependent way. Under optimal conditions the synthetic nonmetabolizable androgen 17 beta-hydroxy-17 alpha-methyl-(3H)-estra-4,9,11-trien-3-one (R1881) (0.1 nM) stimulated cell growth by approximately 2.3 times. Increasing doses of R1881 (1-100 nM) partly decreased the stimulation of the cell growth. The anti-androgen cyproterone acetate exerted inhibitory effects on cell growth. The nuclear extract of the LNCaP-FGC cells contained 17,000 +/- 2,500 (mean +/- SD) KCl-extractable, nuclear androgen receptor sites/cell. Estrogen and progesterone receptors were not detectable in the nuclear extracts nor in cytosol, indicating that these receptors were absent. The release of proteins in the culture medium was studied using incorporation of the 35S-methionine, sodium dodecyl sulfate (SDS)-gel electrophoresis, and fluorography. Cells grown in media containing charcoal-stripped fetal calf serum released significantly lower amounts of a protein with an apparent molecular mass of 42,000 daltons. The release of this 42-kD protein could be restored in cells cultured in the presence of 5 alpha-dihydrotestosterone (0.1-1 microM) or R1881 (0.1-100 nM), whereas the addition of estrogens or corticosteroids had no effect. In the presence of anti-androgens, such as cyproterone acetate and 5,5-dimethyl-3-(4-nitro-3-(trifluoro-methyl)-phenyl)-2,4-imidasolidin edione, inhibitory effects on the release of the 42-kD protein were observed. The observed parallel between the effects of (anti)-androgens on the growth of the LNCaP prostate cells and the release of the 42-kD protein suggests that this protein is involved in the regulation of malignant prostate cell growth.
已对激素敏感性进行了研究，其指标包括类固醇激素受体的存在，以及雄激素和抗雄激素对人前列腺肿瘤细胞系前列腺淋巴结癌快速生长集落（LNCaP-FGC）培养细胞蛋白质释放的影响。LNCaP-FGC细胞的生长受到雄激素的剂量依赖性刺激。在最佳条件下，合成的不可代谢雄激素17β-羟基-17α-甲基-（3H）-雌甾-4,9,11-三烯-3-酮（R1881）（0.1 nM）可使细胞生长刺激约2.3倍。R1881剂量增加（1-100 nM）会部分降低对细胞生长的刺激。抗雄激素醋酸环丙孕酮对细胞生长有抑制作用。LNCaP-FGC细胞的核提取物含有17,000±2,500（平均值±标准差）个可被KCl提取的核雄激素受体位点/细胞。在核提取物和胞质溶胶中均未检测到雌激素和孕激素受体，表明这些受体不存在。使用35S-甲硫氨酸掺入、十二烷基硫酸钠（SDS）-凝胶电泳和荧光自显影法研究了培养基中蛋白质的释放。在含有经活性炭处理的胎牛血清的培养基中生长的细胞释放的一种表观分子量为42,000道尔顿的蛋白质明显较少。在5α-二氢睾酮（0.1-1 microM）或R1881（0.1-100 nM）存在的情况下培养的细胞中，这种42-kD蛋白质的释放可恢复，而添加雌激素或皮质类固醇则无作用。在抗雄激素如醋酸环丙孕酮和5,5-二甲基-3-（4-硝基-3-（三氟甲基）-苯基）-2,4-咪唑烷二酮存在的情况下，观察到对42-kD蛋白质释放的抑制作用。观察到（抗）雄激素对LNCaP前列腺细胞生长和42-kD蛋白质释放的影响之间存在平行关系，这表明该蛋白质参与了恶性前列腺细胞生长的调节。
