前列腺癌為男性最常見的惡性腫瘤之一，並且在西方國家中其為男性癌症排名第三的致命惡性腫瘤。而在台灣近10年來，前列腺癌的發生率與死亡率也都有逐年上升的趨勢。目前用於治療前列腺癌的方式主要是透過遏止雄性素的產生，而這種方式對於早期前列腺癌是具有良好的治療效果。然而，許多病人前列腺癌的惡化，可能肇因於長期使用抗雄性素的療法而使其復發並發展成為一種快速增生及易轉移的雄性素非依賴型前列腺癌。已知，大量活化轉錄因子 CCAAT/enhancer binding protein delta (CEBPD) 會誘導癌細胞的生長停滯以及癌細胞凋亡，因此被視為一抑癌基因。在本篇研究分析發現，CEBPD 能夠被雄性素活化，因此推測 CEBPD 可能在前列腺癌的發展進程扮演關鍵的角色。然而，我們也發現當前列腺癌轉變為雄性素非依賴型態的過程中，E2F1、EZH2 及 SUZ12 的增加會造成 CEBPD 基因的靜默。進一步地，我們發現細胞凋亡啟動者 ─ 蛋白水解酵素Caspase 8 為 CEBPD的下游活化基因，利用 reporter assay 及 in vivo DNA binding assay 證實 CEBPD會直接結合至Caspase 8 的啟動子上並促進其轉錄活化。因此，以“重新活化 CEBPD 的功能”來毒殺前列腺癌為概念，並且跳脫雄性素非依賴型前列腺癌中 CEBPD 啟動子上的外顯遺傳修飾作用，我們建立一種前驅藥物 ─ 特定蛋白酶 perforin 融合蛋白的系統用以治療雄性素依賴型及雄性素非依賴型前列腺癌。實驗結果證實，合併使用 CEBPD 前驅藥物 (提高 procaspase 8 的表現) 和前驅藥物 Granzyme B (活化 procaspase 8 ) 可大量誘發前列腺癌細胞凋亡活性並抑制其生長。因此，此一特定蛋白酶perforin 融合蛋白前驅藥物的應用，可為前列腺癌提供新穎的的治療方法。

Prostate cancer is the most common malignancy in men and the third leading cause of male cancer-related deaths in the Western world. In Taiwan, the incidence of prostate cancer has been rapidly increasing in the past 10 years. The standard therapies include androgen ablation that initially causes tumor regression. However, tumor cells will eventually relapse and develop into castration-resistant prostate cancer (CRPC). Overexpression of transcription factor CCAAT/enhancer binding protein delta (CEBPD) takes part in inducing growth arrest and apoptosis in cancer cells, which could serve as a potential tumor suppressor. Notably, in this study we demonstrated CEBPD is an androgen responsive gene, suggesting CEBPD may play a functional role in progression and development of prostate cancer. Moreover, we found that the activation of E2F1/EZH2/SUZ12 axis participates in CEBPD silencing in CRPC. We next identified that Caspase 8 gene, an apoptosis activator, is a CEBPD downstream target. Reporter assay and in vivo DNA binding assay revealed that CEBPD directly binds and activates Caspase 8 promoter activity. To achieve the idea of “reactivation of CEBPD” for the therapy of prostate cancer and to avoid the epigenetic effects on CEBPD promoter in CRPC, we therefore applied a protease-specific perforin-fused prodrug system for this issue. Our results suggest that the combination of CEBPD prodrug (induction of procaspase 8 level) and granzyme B prodrug (an activator of procaspase 8) could trigger stronger apoptotic effect and impede their proliferation in androgen dependent prostate cancer and CRPC. Accordingly, protease-specific perforin-fused prodrug therapy offers a new therapeutic strategy in treating prostate cancers.

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