Identification of phosphorylated proteins involved in the oncogenesis of prostate cancer via Pin1-proteomic analysis
Identification of phosphorylated proteins involved in the oncogenesis of prostate cancer via Pin1-proteomic analysis
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DOI：
10.1002/pros.21466
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发表时间：
2012-05-01
期刊：
影响因子：
2.8
通讯作者：
Ryo, Akihide
中科院分区：
文献类型：
--
作者：
Endoh, Kanji;Nishi, Mayuko;Ryo, Akihide
肽基脯氨酰异构酶Pin1通过催化特定磷酸化Ser/Thr-Pro基序的顺反异构化来调节磷酸化蛋白质的亚类。虽然Pin1已被证明参与细胞转化和前列腺癌恶性表型的维持，但其在这些过程中的特异性底物尚未确定。通过用重组GST-Pin1蛋白进行GST-pull down分析，从两种人前列腺癌细胞系（PC-3，LNCaP）和Dunning大鼠前列腺癌细胞系中分离癌症特异性磷酸化蛋白。然后使用Q-Tof微型质谱仪通过LC-MS/MS分析鉴定这些蛋白质，并进行进一步的功能分析。我们新发现了五个前列腺癌特异性Pin1结合蛋白（PINBPs）。其中，TRK融合基因（TFG）被发现在前列腺癌细胞系和组织中优先上调。通过特异性siRNA靶向抑制TFG导致PC3前列腺癌细胞中细胞增殖减少和诱导早衰。我们进一步发现TFG可以促进NF-κ B和雄激素受体（AR）介导的细胞信号传导。基于组织显微切割的定量RT-PCR分析前列腺癌根治性切除术后的组织进一步显示TFG表达与手术后肿瘤复发的可能性更高和时间更短密切相关。基于Pin1的蛋白质组学分析是鉴定前列腺癌特异性磷酸化蛋白的有用工具。TFG可能是前列腺癌潜在的诊断和/或预后标志物和治疗靶点。前列腺72：626 - 637，2012年。(C)2011 Wiley Periodicals，Inc.
BACKGROUND The peptidyl-prolyl isomerase Pin1 regulates a subset of phosphorylated proteins by catalyzing the cis-trans isomerization of their specific phosphorylated Ser/Thr-Pro motifs. Although Pin1 has been shown to be involved in cell transformation and the maintenance of the malignant phenotype in prostate cancer, its specific substrates during these processes have not yet been determined.METHODS. Cancer-specific phosphorylated proteins were isolated from two human prostate cancer cell lines (PC-3, LNCaP) and the Dunning rat prostate cancer cell lines by GST-pull down analysis with recombinant GST-Pin1 protein. These proteins were then identified by the LC-MS/MS analysis using a Q-Tof micro mass spectrometer and processed for further functional analysis.RESULTS. We newly identified five prostate cancer-specific Pin1 binding proteins (PINBPs) in this screen. Among these, TRK-fused gene (TFG) was found to be preferentially up-regulated in prostate cancer cell lines and tissues. The targeted inhibition of TFG by specific siRNA resulted in the reduced cell proliferation and the induction of premature senescence in PC3 prostate cancer cells. We further found that TFG can facilitate the cell signaling mediated by NF-kappaB and androgen receptor (AR). Tissue micro-dissection based quantitative RT-PCR analysis of prostate cancer tissues following radical prostatectomy further revealed that TFG expression is closely associated with both a higher probability and shorter period of tumor recurrence following surgery.CONCLUSIONS. Pin1-based proteomics analysis is a useful tool for the identification of prostate cancer-specific phosphorylated proteins. TFG could be a potential diagnostic and/or prognostic marker and therapeutic target in prostate cancer. Prostate 72:626-637, 2012. (C) 2011 Wiley Periodicals, Inc.