多形性神經膠質母細胞瘤(Glioblastoma multiforme, GBM)是一種非常具有侵略性而且相當惡性末期的腦神經膠質瘤(glioma)。在歐洲及北美地區，每年每十萬人中就有2~3人被診斷出罹患GBM，平均在診斷出罹患GBM後病人的壽命通常只剩下約兩年的時間，即使利用手術切除腫瘤再加上放療、化療藥物的輔助治療，病人大多數仍會死於極高的復發率，所以目前最迫切的課題在於找出GBM為何如此具有抵抗性和其再復發性，好讓我們能更有效地去了解癌症形成的原因和建立其致病模式。Hippo pathway在哺乳類中扮演著控制器官大小的關鍵角色，藉由調控細胞的增生與凋亡來達成。當Hippo pathway中的MST1/2失去活性時，Hippo pathway最主要的下游調控蛋白YAP1會大量表現，從細胞質進入細胞核中和TEAD轉錄因子結合而活化許多和細胞增生與抑制細胞凋亡相關基因的表現，YAP1大量表現容易造成器官的過度生長而有癌化的現象發生，所以Hippo pathway也被稱作是一種抑癌基因路徑。根據一些研究指出，我們發現在許多的癌症中YAP1經常是扮演著致癌基因、具有高度表現量的角色，所以本篇的研究重點首先在於利用實驗室先前建立的老鼠glioma和GBM細胞株探討MST1和YAP1在腦癌中扮演之角色並尋找Hippo pathway與腦癌疾病致病分子路徑的交互作用為何?此外在腦癌的治療上，我們也找出YAP1 inhibitor-Verteporfin (VP)確實能對腦癌細胞株造成細胞凋亡的效果。最後我們也利用了Mst1/2缺失造成的YAP1的大量表現並搭配抑癌基因p53的缺失，兩者特異性地缺失在腦部的星狀膠質細胞，利用基因鼠動物模式，觀察MST1/2對於glioma或者是GBM惡化的影響機制為何，希望能建立一套Hippo pathway與GBM活體動物模式的研究平台並從中找出一些重要的GBM治療標靶節點。

Glioblastoma multiforme (GBM) is the most aggressive and malignant primary brain tumor. Primary glioblastomas have a worse prognosis with a median survival of less than 2 years after diagnosis. The Hippo signaling transduction pathway plays multiple functions during organ development, adult tissues homeostasis and tumorigenesis, and its downstream mediator yes-associated protein 1 (YAP1) is known to function as a potent oncogene, which is amplified and overexpressed in various human cancers. The goal of this study is to study the role of Hippo pathway in GBM carcinogenesis. First, we selected primary murine glioblastoma cancer cell lines as an in vitro model system to study the functional roles of Hippo pathway in regulating the proliferation, adhesion and migration of GBM cells in vitro. Second, to identify the role of Hippo pathway in brain development, we selected GFAP drive Cre transgenic system to cross with conditional Mst1/Mst2 loxp mice to specific knockout Mst1/Mst2 (upstream kinases of Hippo pathway) expression in the glial cells during brain development. Moreover, to discovery the important function of the Hippo signaling involved in the development of GBM, we have established mouse models of GBM (activation of KrasG12D combined with p53 loss) for studying the critical roles of Hippo pathway in GBM progression. An understanding of the temporal regulation of Hippo during GBM progression and the identification of activation Hippo signaling combined p53 induced a transcriptional profiles— with respect to upstream activators and downstream networks, which may play an important node in the design of clinical approaches targeting Hippo signaling for the treatment of GBM.

論文審定書 i
Abstract in Chinese ii
Abstract in English iii
Abbreviations v
Introduction 1
Materials and Methods 9
Results 20
Discussion 29
Figures and Tables 36
References 71

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