Secretion of prostatic specific antigen, proliferative activity and androgen response in epithelial-stromal co-cultures from human prostate carcinoma
Secretion of prostatic specific antigen, proliferative activity and androgen response in epithelial-stromal co-cultures from human prostate carcinoma
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DOI：
10.1111/j.1365-2605.2004.00509.x
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发表时间：
2005-02-01
影响因子：
--
通讯作者：
Huidobro, C
中科院分区：
文献类型：
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作者：
Castellón, E;Venegas, K;Huidobro, C
我们调查的增殖活性，前列腺特异性抗原（PSA）的分泌，形态和雄激素反应的人前列腺肿瘤上皮细胞与基质细胞共培养在一个双室系统。采用酶消化法分离前列腺腺癌基质细胞和上皮细胞，并在限定培养基中培养。进行前列腺癌肿瘤抗原（PCTA-1）的免疫细胞化学以评价培养物纯度。此外，共培养的上皮细胞的形态进行了评价，通过电子显微镜。PSA通过微粒酶免疫测定（MEIA）自动化方案测定，增殖通过商业分光光度试剂盒评估，基于甲瓒盐的形成。两种细胞培养物均显示纯度超过90%。上皮细胞共培养物显示出明显的膜突起和细胞交错。基质细胞的存在下，上皮细胞的增殖活性增加。此外，PSA分泌显著增加并维持至少14天，而PSA分泌的雄激素反应仅在共培养条件下得到证实。人前列腺癌上皮细胞和间质细胞的原代共培养物能够在较长时间内维持增殖和分泌特性以及激素反应，并且代表前列腺癌细胞和分子研究的有价值的工具。
We investigate the proliferative activity, prostatic specific antigen (PSA) secretion, morphology and androgen response of human prostate tumour epithelial cells co-cultured with stromal cells in a bicameral system. Stromal and epithelial cells were isolated from prostate adenocarcinoma by enzyme digestion and cultured in defined media. Immunocytochemistry for prostate carcinoma tumour antigen (PCTA-1) was performed for culture purity evaluation. Also, the morphology of the epithelial cells in co-culture was evaluated by electron microscopy. PSA was determined by microparticle enzyme immunoassay (MEIA) automatized protocol and the proliferation was evaluated by a commercial spectrophotometric kit, based on formazan salt formation. Both cell cultures showed more than 90% of purity. The epithelial cell co-cultures showed marked membrane processes and cell interdigitations. The proliferative activity of the epithelial cells was increased in presence of stromal cells. Also, PSA secretion was significantly increased and maintained for at least 14 days, whereas the androgen response for PSA secretion was evidenced only in co-culture condition. Primary co-cultures of epithelial and stromal cells from human prostate carcinoma are able to maintain, for a prolonged time, proliferative and secretory properties as well hormone response, and represent a valuable tool for cellular and molecular studies on prostate cancer.