Collins A T, Zhiming B, Gilmore K, Neal D E
Department of Surgery, Medical School, University of Newcastle, Newcastle upon Tyne, UK.
J Endocrinol. 1994 Nov;143(2):269-77. doi: 10.1677/joe.0.1430269.
Stromal cells derived from collagenase-digested benign hyperplastic adult prostates were isolated and grown in culture. Androgen and oestrogen receptor status were determined and growth in response to mibolerone (a synthetic androgen) and oestradiol-17 beta was measured. In addition, the ability of oestrogens to regulate the androgen receptor in stromal cells was investigated. [3H]Thymidine incorporation into DNA was stimulated by mibolerone in primary and secondary cultures, but sensitivity was lost with subsequent passages. Androgen stimulation of [3H]thymidine incorporation was consistently inhibited by the anti-androgen cyproterone acetate. Oestradiol-17 beta also stimulated [3H]thymidine incorporation into DNA, and this effect was inhibited by the anti-oestrogen tamoxifen. Sensitivity to oestradiol was lost with subsequent passages. A combination of mibolerone and oestradiol was not synergistic in increasing [3H]thymidine incorporation into DNA, but maximal stimulation occurred at 100-fold lower concentrations of mibolerone and oestradiol when the two hormones were applied in combination. Specific high-affinity [3H]mibolerone- and [3H]oestradiol-binding sites were demonstrated by radioligand binding in intact cells. The affinity for oestradiol binding to its receptor exceeded that quantified for mibolerone binding to the androgen receptor, whilst the number of oestradiol-binding sites was approximately tenfold less than that quantified for mibolerone. Treatment with oestradiol down-regulated the number of [3H]mibolerone binding sites 1.7-fold (P < 0.005) as early as day 2 after oestradiol treatment. In conclusion, we successfully cultured stromal cells derived from hyperplastic prostates which retained sensitivity to androgen and oestrogen.(ABSTRACT TRUNCATED AT 250 WORDS)
从经胶原酶消化的良性增生成年前列腺中分离出基质细胞，并在培养中生长。测定雄激素和雌激素受体状态，并检测其对米勃酮（一种合成雄激素）和雌二醇-17β的反应性生长。此外，还研究了雌激素调节基质细胞中雄激素受体的能力。在原代和传代培养中，米勃酮刺激[3H]胸腺嘧啶核苷掺入DNA，但随着传代次数增加敏感性丧失。抗雄激素醋酸环丙孕酮持续抑制雄激素对[3H]胸腺嘧啶核苷掺入的刺激作用。雌二醇-17β也刺激[3H]胸腺嘧啶核苷掺入DNA，且这种作用被抗雌激素他莫昔芬抑制。随着传代次数增加对雌二醇的敏感性丧失。米勃酮和雌二醇联合使用在增加[3H]胸腺嘧啶核苷掺入DNA方面无协同作用，但当两种激素联合应用时，在米勃酮和雌二醇浓度低100倍时出现最大刺激作用。通过放射性配体结合在完整细胞中证明了特异性高亲和力的[3H]米勃酮和[3H]雌二醇结合位点。雌二醇与其受体结合的亲和力超过米勃酮与雄激素受体结合的定量亲和力，而雌二醇结合位点的数量比米勃酮定量的数量少约10倍。早在雌二醇处理后第2天，雌二醇处理使[3H]米勃酮结合位点数量下调1.7倍（P<0.005）。总之，我们成功培养了源自增生前列腺的基质细胞，这些细胞对雄激素和雌激素仍保持敏感性。（摘要截短于250字）
