Transdifferentiation of cultured human prostate cancer cells to a neuroendocrine cell phenotype in a hormone-depleted medium.
Transdifferentiation of cultured human prostate cancer cells to a neuroendocrine cell phenotype in a hormone-depleted medium.
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DOI：
10.1016/s1078-1439(97)00039-2
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发表时间：
1997-03-01
期刊：
影响因子：
--
通讯作者：
Buttyan, R
中科院分区：
文献类型：
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作者：
Shen, R;Dorai, T;Buttyan, R
神经内分泌（NE）细胞被神秘地发现与人类前列腺癌有关，据报道，它们的数量在晚期和耐药肿瘤中增加。这种细胞类型的起源和它们在前列腺肿瘤中出现的原因仍然没有解决。先前，Bang等人（Proc Natl Acad Sci USA 1994;91：5330）报道了二丁酰腺苷3 ′，5 ′-环磷酸（db-cAMP），一种上调细胞内cAMP的试剂，能够诱导培养的人前列腺癌细胞（包括雄激素敏感性LNCaP系）的NE细胞样表型。在这里，我们报告说，慢性孵育LNCaP细胞在培养基中含有10%的木炭剥离胎牛血清（CSFBS）同样诱导NE分化这些细胞。在将LNCaP细胞的低密度培养物转换到这种改良培养基的5天内，细胞生长停滞并获得改变的形态，具有许多细胞质分泌颗粒和类似于培养的神经元的伸长的突起。这种形态在10天时占优势，在培养20天时观察到完全转化。CS-FBS维持的细胞切片的电子显微镜分析显示存在大量NE细胞特征性的致密核心分泌颗粒。免疫组织化学染色确定了NE标记蛙皮素，神经元特异性烯醇化酶，和S-100在这种改良的培养基中的表达上调。一旦建立，发现NE细胞样表型在用含有未修饰的胎牛血清的培养基替换时是可逆的，但通过直接补充具有二氢睾酮（DHT）（InM）的CS-FBS培养基则不是。然而，补充DHT确实抑制了NE细胞样表型的发展，当它存在于暴露于CS-FBS培养基的开始时。相反，db-cAMP治疗，这并不影响前列腺特异性抗原（PSA）或雄激素受体（AR）的LNCaP细胞的表达，NE分化的LNCaP细胞来源于这种前列腺素缺乏的培养基显示显着下调PSA和AR的表达。这些体外结果进一步支持了前列腺癌细胞可以在体内转化为具有NE表型的细胞的概念，并表明这种转化可能通过前列腺癌的某些疗法在患者中加速。
Neuroendocrine (NE) cells are enigmatically found in association with human prostate cancers and their numbers are reported to increase in advanced and hormoneresistant tumors. The origin of this cell type and the reason for their appearance in prostate tumors remains unresolved. Previously, Bang et al. (Proc Natl Acad Sci USA 1994;91:5330) reported that dibutyryl adenosine 3',5'-cyclic phosphate (db-cAMP), an agent that upregulates intracellular cAMP, was able to induce a NE cell-like phenotype of cultured human prostate cancer cells, including the androgen-sensitive LNCaP line. Here we report that chronic incubation of LNCaP cells in a medium containing 10% charcoal-stripped fetal bovine serum (CSFBS) likewise induces NE differentiation of these cells. Within 5 days of switching low density cultures of LNCaP cells to this modified medium, the cells growth arrest and acquire an altered morphology with numerous cytoplasmic secretory granules and elongated processes that resemble cultured neurons. This morphology predominates at 10 days with complete transformation seen by 20 days of culture. Electron microscopic analysis of sections of CS-FBS maintained cells showed the presence of abundant dense core secretory granules characteristic of NE cells. Immunohistochemical staining identified the upregulation of the expression of NE markers bombesin, neuron-specific enolase, and S-100 in this modified culture medium. Once established, the NE cell-like phenotype was found to be reversible upon replacement with a medium containing unmodified fetal bovine serum, but not by direct supplementation of CS-FBS medium with dihydrotestosterone (DHT) (I nM). DHT supplementation did, however, suppress the development of the NE cell-like phenotype when it was present at the initiation of exposure to CS-FBS medium. In contrast to db-cAMP treatment, which did not affect prostate specific antigen (PSA) or androgen receptor (AR) expression of LNCaP cells, NE-differentiated LNCaP cells derived in this hormone-deficient medium showed marked downregulation of PSA and AR expression. These in vitro results further support the concept that prostate cancer cells can tranform in vivo to cells with a NE phenotype and suggest that this transformation might be accelerated in patients by certain therapies for prostate cancer.