Anterior gradient 2 and 3 - two prototype androgen-responsive genes transcriptionally upregulated by androgens and by oestrogens in prostate cancer cells
Anterior gradient 2 and 3 - two prototype androgen-responsive genes transcriptionally upregulated by androgens and by oestrogens in prostate cancer cells
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DOI：
10.1111/febs.12118
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发表时间：
2013-03-01
期刊：
影响因子：
5.4
通讯作者：
Klocker, Helmut
中科院分区：
文献类型：
--
作者：
Bu, Huajie;Schweiger, Michal R.;Klocker, Helmut
雄激素和雌激素与前列腺癌发生和肿瘤进展有关。虽然雄激素的作用已被广泛研究，但前列腺癌中雌激素信号传导的机制尚未完全了解。在本研究中，我们分析了雄激素和雌激素对前梯度2（AGR 2）和前梯度3（AGR 3）表达的影响，前梯度2和前梯度3包括两个高度相关的基因，编码在前列腺癌中表达的分泌蛋白，其中之一（AGR 2）与肿瘤转移相关。定量逆转录酶PCR和蛋白质印迹分析表明，在三个雄激素受体阳性细胞系中，AGR2和AGR3的雄激素诱导，从0.1nm的浓度开始。这两个AGR基因也转录激活5nm雌二醇，但不是同种型选择性或非选择性雌激素受体激动剂在DUCaP细胞，港口的野生型雄激素受体的高水平。雄激素和雌激素调节都需要一个功能性雄激素受体，而不是雌激素受体。这种雄激素和雌激素调节的模式在VCaP细胞中得到证实，并且也在FKBP5中观察到，FKBP5是一种充分表征的雄激素调节基因。全基因组染色质免疫沉淀研究结合深度测序鉴定了分别位于AGR2和AGR3基因远端启动子和内含子区域的雄激素受体结合位点。这些增强子的雄激素反应性通过荧光素酶报告基因测定和定点突变分析来验证。雄激素治疗还诱导p300和RNA PolII募集到AGR2的雄激素受体增强子，并启动局部染色质重塑和含RNA PolII的雄激素受体转录复合物的形成。
Androgens and oestrogens have been implicated in prostatic carcinogenesis and tumour progression. Although the actions of androgens have been studied extensively, the mechanisms underlying oestrogen signalling in prostate cancer are not fully understood. In the present study, we analyzed the effect of androgens and oestrogens on the expression of anterior gradient2 (AGR2) and anterior gradient3 (AGR3), comprising two highly-related genes encoding secretory proteins that are expressed in prostate cancer and one of which (AGR2) has been associated with tumour metastasis. Quantitative reverse-transcriptase PCR and western blot analysis showed androgen induction of AGR2 and AGR3 in three androgen receptor positive cell lines, starting at concentrations of 0.1nm. Both AGR genes were also transcriptionally activated by 5nm oestradiol but not by isotype selective or nonselective oestrogen receptor agonists in DUCaP cells that harbour a high-level of wild-type androgen receptor. A functional androgen receptor but not oestrogen receptor turned out to be required for both androgen and oestrogen regulation. This pattern of androgen and oestrogen regulation was confirmed in VCaP cells and was also observed for FKBP5, a well-characterized androgen-regulated gene. Genome-wide chromatin-immunoprecipitation studies coupled with deep sequencing identified androgen receptor binding sites localized in the distal promoter and intron regions of the AGR2 and AGR3 genes, respectively. The androgen responsiveness of these enhancers was verified by luciferase reporter gene assays and site-directed mutagenesis analysis. Androgen treatment also induced p300 and RNA PolII recruitment to androgen receptor enhancers of AGR2 and initiated local chromatin remodelling and the formation of RNA PolII-containing androgen receptor transcription complexes.