胃癌是一個進展快且具有高致死率的癌症，特別好發於亞洲地區。在分子病
理研究中，胃癌的起始及發展和 KRAS 的活化、E-cadherin 蛋白減少、P53、APC
及 PTEN 的失活有關，其相關機制都是在研究胃癌腫瘤的重要基礎。LKB1 為
serine/threonine 蛋白激酶，主要是腺嘌呤磷酸激酶活化蛋白激酶(AMPK)的上游活
化激酶，並且調控 E-cadherin 蛋白的表現。在過去的研究中，LKB1 的突變會導
致 Peutz-Jeghers 症候群，病人在胃腸道中會有息肉及惡性腫瘤的發生。PTEN 為
一種酪氨酸磷酸酶，其調節 AKT 途徑的活化，經常突變在許多類型的人類癌
症。
本研究利用小鼠之動物模式，搭配我們實驗室開發之 H + /K + ATPase Ccre，特
異性在在胃組織中同時剔除 LKB1 及 PTEN 基因，誘導使小鼠誘發產生和人類相
似之胃腺癌。而在剃除 LKB1 及 PTEN 的小鼠疾病模型中，發現小鼠外觀有出現
貧血之現象，以血液生化檢驗後更發現白血球大量升高，並且伴隨血小板增高。
在糞便也偵測到有潛血反應。在組織切片中確認更發現小鼠胃腫瘤組織呈現胃腺
癌之組織型態，並且在免疫組織染色中發現 LKB1 下游磷酸化 AMPK 減少、
PTEN 下游之磷酸化 AKT/mTOR 增加，及腫瘤幹細胞標記 CD44 上升、E-
cadherin 表現下降以及上皮-間質轉換之標記 SMA、MMP9 等上升，此外，也在
胃癌組織中體外培養出初代胃癌細胞，證明此基因轉殖鼠疾病模型具有能產生惡
性胃腺癌之能力。最後，人類胃腺癌細胞 AGS 剃除 LKB1 的表現，發現細胞生
長速度、癒合能力增加，與癌細胞之體外行為表現與活體內腫瘤進展行為不謀而
合。希望藉由此動物模式追加分析探討其組織病變分子改變模式、血管新生、腫
瘤微環境之改變，此研究結果將有助於了解並解釋胃炎臨床之基因病變與徵狀的
關係發生，以及潛在可能惡化的模式因素，全面性了解胃癌惡化的機轉、並開發
新的治療打擊胃癌的相關藥物。

Gastric cancer (GC) is an aggressive disease with the highest rate of mortality among cancers and is the second leading cause of cancer death worldwide. Recent molecular pathology studies have elucidated a detailed profile of genetic lesions associated with GC initiation and progression—activation KRAS and inactivation or loss of E-cadherin, P53, APC and PTEN—providing a foundation for investigation of the biological and biochemical basis for this malignancy. LKB1, also known as Serine/threonine kinase 11 (STK11), encodes a serine/threonine kinase that directly phosphorylates and activates AMPK, a cellular metabolic kinase. LKB1-AMPK pathway function also includes the maintenance of epithelial junction stability by regulating E-cadherin expression. In addition, LKB1 mutations are associated with the Peutz-Jeghers syndrome (PJS) consisting of intestinal polyposis and other gastrointestinal malignancies. PTEN, a tyrosine phosphatase, which regulates the activation of AKT pathway, is frequently mutated in many types of human cancer. This study seeks to use the H+/K+ ATPase Cre transgene mice, developed in our lab, directs Cre recombinase to the stomach commencing to both abrogate LKB1 and PTEN function in a stomach-specific manor to induce the development and metastatic progression of gastric adenocarcinomas, with similarities to human gastric adenocarcinoma. We also determine how these alterations contributes to gastric tumor histopathologic progression, invasive and metastatic potential, angiogenic and tumor stromal microenvironment. Meanwhile, information about responses in a bona fide GC model will likely contribute to interpreting those clinical results, are they positive or negative, and potentially might help guide further trial designs to combat stomach cancer.

國立中山大學研究生學位論文審定書+i
中文摘要+ii
Abstract+iii
Abbreviation+v
Introduction+1
Material and Method+8
Results+16
Discussion+27
Figures+33
Tables+59
References+62

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