Biochemical characterization of nuclear receptors for vitamin D3 and glucocorticoids in prostate stroma cell microenvironment
Biochemical characterization of nuclear receptors for vitamin D3 and glucocorticoids in prostate stroma cell microenvironment
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DOI：
10.1016/j.bbrc.2011.06.181
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发表时间：
2011-08-19
影响因子：
3.1
通讯作者：
Onate, Sergio A.
中科院分区：
文献类型：
--
作者：
Hidalgo, Alejandro A.;Montecinos, Viviana P.;Onate, Sergio A.
前列腺组织微环境中基质细胞信号的破坏影响前列腺癌向雄激素独立发展。1 α，25-二羟基维生素D-3 （1,25 d(3)）和糖皮质激素，单独或联合，已被研究作为治疗雄激素治疗失败的晚期前列腺癌的替代方案。糖皮质激素的作用是由细胞内糖皮质激素受体介导的。同样，1,25 d(3)的作用是由1,25 d(3)核受体（VDR）介导的。本研究从人前列腺中分离出良性（BAS）和癌相关基质（CAS）的成纤维细胞，以表征VDR和GR在前列腺基质中的转录因子功能。使用cyp24 -荧光素酶报告基因评估的vdr介导的转录活性在13个CAS中有9个（70%）被1,25 d(3)限制为3倍诱导，相比之下，在BAS临床样本对中有100个10倍诱导。在9个CAS中的7个中，his标记的VDR （Ad-his-VDR）的表达未能恢复荧光素酶报告基因的低转录活性。有趣的是，在分析的9个CAS中，有2个Ad-his-VDR的表达成功恢复了受体介导的诱导，这表明受体蛋白本身的变化是导致对1,25 d(3)作用的反应和抗性降低的原因。相反，与BAS样本对相比，13个CAS样本对中有4个（30%）的vdr介导的转录活性更有效。与在CAS中观察到的对1,25 d(3)的反应降低一致，染色质免疫沉淀（ChIP）试验显示共激活因子SRC-1/CBP的募集减少，而VDR对CYP24启动子的募集没有重大变化。此外，我们观察到，与BAS相比，CAS中gr介导的转录活性也发生了改变。干扰共激活因子SRC-1/CBP的募集可能会促进CaP中的激素抵抗，并强调了肿瘤细胞微环境中分子诊断和药物设计的相关性。(C) 2011爱思唯尔公司版权所有。
The disruption of stromal cell signals in prostate tissue microenvironment influences the development of prostate cancer to androgen independence. 1 alpha,25-Dihydroxyvitamin D-3 (1,25D(3)) and glucocorticoids, either alone or in combination, have been investigated as alternatives for the treatment of advanced prostate cancers that fails androgen therapies. The effects of glucocorticoids are mediated by the intracellular glucocorticoid receptor (GR). Similarly, the effect of 1,25D(3) is mediated by the 1,25D(3) nuclear receptor (VDR). In this study, fibroblasts from benign- (BAS) and carcinoma-associated stroma (CAS) were isolated from human prostates to characterize VDR and GR function as transcription factors in prostate stroma. The VDR-mediated transcriptional activity assessed using the CYP24-luciferase reporter was limited to 3-fold induction by 1,25D(3) in 9 out of 13 CAS (70%), as compared to >10-fold induction in the BAS clinical sample pair. Expression of His-tagged VDR (Ad-his-VDR) failed to recover the low transcriptional activity of the luciferase reporter in 7 out of 9 CAS. Interestingly, expression of Ad-his-VDR successfully recovered receptor-mediated induction in 2 out of the 9 CAS analyzed, suggesting that changes in the receptor protein itself was responsible for decreased response and resistance to 1,25D(3) action. Conversely, VDR-mediated transcriptional activity was more efficient in 4 out of 13 CAS (30%), as compared to the BAS sample pair. Consistent with the reduced response to 1,25D(3) observed in CAS, chromatin immunoprecipitation (ChIP) assays indicated decreased recruitment of coactivators SRC-1/CBP, without major changes in the recruitment of VDR to the CYP24 promoter. In addition, we observed that GR-mediated transcriptional activity was also altered in CAS, as compared to BAS. Disruption of coactivators SRC-1/CBP recruitment may promote hormone resistance in CaP, and highlights the relevance of molecular diagnosis and drug design in tumor cell microenvironment. (C) 2011 Elsevier Inc. All rights reserved.