TRPM8 channel as a novel molecular target in androgen-regulated prostate cancer cells.
TRPM8 channel as a novel molecular target in androgen-regulated prostate cancer cells.
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DOI：
10.18632/oncotarget.3948
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发表时间：
2015-07-10
期刊：
影响因子：
--
通讯作者：
Zakharian E
中科院分区：
文献类型：
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作者：
Asuthkar S;Velpula KK;Elustondo PA;Demirkhanyan L;Zakharian E
寒冷和薄荷醇受体TRPM8在前列腺和前列腺癌（PC）中高表达。最近，我们发现TRPM8是一种离子型睾酮受体。TRPM8 mRNA在雄激素水平高的早期前列腺肿瘤中表达，而抗雄激素治疗大大降低了其表达。通过染色质免疫沉淀（ChIP）分析，我们发现雄激素反应元件（ARE）介导trpm8的雄激素调节。此外，通过免疫荧光、钙成像和平面脂质双分子层，我们发现前列腺中的TRPM8通道在功能上受到雄激素的调节。尽管TRPM8 mRNA在高水平表达，但我们发现TRPM8蛋白在PC细胞中经历泛素化和降解。LNCaP细胞免疫沉淀的TRPM8质谱分析鉴定出泛素样修饰物激活酶1 （UBA1）。PYR-41是泛素化级联初始酶UBA1的有效抑制剂，可增加LNCaP细胞质膜（PM）上TRPM8的活性。此外，pyr -41介导的PMTRPM8活性伴随着p53和Caspase-9的增强激活。有趣的是，我们发现trpm8启动子具有p53的推定结合位点，并且p53的过表达增加了trpm8 mRNA水平。除了AR和p53对TRPM8的基因组调控外，我们的研究结果表明，睾酮诱导的PMTRPM8活性诱导Ca2+摄取，随后导致凋亡细胞死亡。这些发现支持通过调节PC细胞生长和增殖来挽救PMTRPM8表达作为一种新的治疗应用的策略。
The cold and menthol receptor TRPM8 is highly expressed in prostate and prostate cancer (PC). Recently, we identified that TRPM8 is as an ionotropic testosterone receptor. The TRPM8 mRNA is expressed in early prostate tumors with high androgen levels, while anti-androgen therapy greatly reduces its expression. Here, from the chromatin-immunoprecipitation (ChIP) analysis, we found that an androgen response element (ARE) mediates androgen regulation of trpm8. Furthermore, using immunofluorescence, calcium-imaging and planar lipid bilayers, we identified that TRPM8 channel is functionally regulated by androgens in the prostate. Although TRPM8 mRNA is expressed at high levels, we found that the TRPM8 protein undergoes ubiquitination and degradation in PC cells. The mass-spectrometry analysis of TRPM8, immunoprecipitated from LNCaP cells identified ubiquitin-like modifier-activating enzyme 1 (UBA1). PYR-41, a potent inhibitor of initial enzyme in the ubiquitination cascade, UBA1, increased TRPM8 activity on the plasma membrane (PM) of LNCaP cells. Furthermore, PYR-41-mediated PMTRPM8 activity was accompanied by enhanced activation of p53 and Caspase-9. Interestingly, we found that the trpm8 promoter possesses putative binding sites for p53 and that the overexpression of p53 increased the TRPM8 mRNA levels. In addition to the genomic regulation of TRPM8 by AR and p53, our findings indicate that the testosterone-induced PMTRPM8 activity elicits Ca2+ uptake, subsequently causing apoptotic cell death. These findings support the strategy of rescuing PMTRPM8 expression as a new therapeutic application through the regulation of PC cell growth and proliferation.