在西方國家中，前列腺癌是男性最常見的癌症。其進展由早期的良性增生形成前列腺上皮腫瘤，隨後進一步發展為惡性腫瘤以及晚期癌細胞轉移到身體其他部位。在先前的文獻中指出，P53 基因的突變主要都是發現在癌症的晚期及轉移的病例中，但對於癌症早期或是原發腫瘤的影響仍然未知；而在最近的文獻中指出，前列腺癌的原發及轉移的病理中發現到RAS/MAPK訊息傳遞路徑的活化扮演非常重要的角色。本研究的目的為建立兩種不同基因工程小鼠來探討P53基因的剃除搭配K-rasG12D或是BRAFV600E突變的活化下對於前列腺癌的發展影響。我們發現這兩種動物模型在隨著時間的進展下，展現出高度的增生能力引發腫瘤形成並造成癌細胞的轉移。另外，在搭配前列腺腫瘤組織分離出的老鼠初代前列腺癌細胞下，結合了免疫組織化學染色及西方墨點法的實驗結果，我們觀察到MAPK訊息路徑具有高度的活化。特別的是，K-rasG12D蛋白還調控活化了另外一條Akt/mTOR 訊息傳遞，進一步比較出K-rasG12D與BRAFV600E這兩者之間所調控的分子機制。接著，透過收集前列腺癌小鼠的血清，我們找出了具有潛力的生物標記，mGluR1。當使用了抑制劑Riluzole處理在前列腺初代癌細胞中，能夠降低細胞增殖/存活表現。西方墨點法分析結果也發現了caspase 3活化，證實mGluR1在前列腺癌細胞的重要性。最後，我們成功的建立出新的前列腺癌基因轉殖動物模型，在未來提供一個非常好的工具來找尋新的抗癌藥物，並應用在前列腺癌的治療。

Prostate cancer (PCa) is one of the most common malignant tumors in men and the second leading cause of cancer deaths in male worldwide. P53 tumor suppressor gene is one of the most frequently inactivated genes in advanced prostate lesions, suggesting that aberrations of p53 would be required for PCa progression. On the other hand, activation of the the RAS/MAPK pathway occurred in both primary and metastatic prostate lesions, whereas p-ERK has been shown to regulate androgen signaling by phosphorylating androgen receptor (AR) coactivators, which may play an important role in prostate carcinogenesis. In this study, we developed two genetically engineered mouse (GEM) models that crossed both conditional activate LSL/KrasG12D or mutant BrafV600E and conditional abrogate p53 function in a prostate-specific manor, recapitulating the PCa histopathology, precursor lesions, and clinical behavior of the human disease with high fidelity. These tissue-specific compound mutant mice develop prostate tumors and metastasis. We also generated primary PCa cell lines derived from two mouse models to characterize and compare their different malignant behaviors in vitro. The phenotypic comparison of the Pb-Cre; K-rasG12D; P53L/L mice with the Pb-cre; BrafV600E; P53L/L mice by immunohistochemistry stain (IHC) and western blot analysis showed that not only MAPK pathway, K-ras also activates the Akt-mTOR signaling. Through proteomic analysis of mouse plasma samples, finding the new biomarker in PCa, mGluR1. Inhibition of mGluR1 by Riluzole in primary PCa cells had reduced cell proliferation/viability and increased caspase 3 activity. Moreover, western blot data found that Riluzole suppress signaling through the MAPK and AKT/mTOR pathways. Finally, our PCa models provide a unique tool for searching potential biomarkers and for drug discovery affecting K-ras signaling can be valuable for treating this disease. Anti-mGluR1 may be contributing a new treatment for prostate cancer.

論文審定書……………………………………………i
Abstract in Chinese……………………………………ii
Abstract in English……………………………...….…iii
Abbreviation……………………………………………v
Contents....................................................vi
Figures and Legends.....................................vii
Introduction………………………………………….... 1
Materials and Methods………………………………...8
Results…………………………………………………….16
Discussion…………………………………………………25
Figures and Tables………………………………………. 29
References……………………………………………….. 47

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