人類從古時候開始就運用了許多陸生動物作為醫藥封面的資源，但到目前為止還是有許多疾病仍無法被根治，由於海洋生物生存環境特殊，需要具有適應高壓、高鹽、低氧的棲地，因此其天然物具有特殊的化學結構。雖然人類運用海洋資源進行藥物篩選的時間比陸地上少了很多，但海洋藥物的成功案例卻比陸地資源高很多，現在已經有許多海洋藥物運用在臨床醫學上，相信不久後海洋的資源可以解決人類現今無法解決的疾病。
由於肝癌是現在台灣癌症十大死亡原因之一且到目前為止，肝癌唯一的標靶藥物sorafenib經過長期觀察，對醫治肝癌的能力與肝癌患者的存活率效果並不顯著，因此開發肝癌新藥是現今重要的課題。本篇研究是從海洋細菌所萃取出的次級代謝產物，並評估其抑制肝癌細胞生長的能力。
先前研究已經發現這種海洋二次代謝產物在大腸癌針對p53突變有明顯的療效並提升p73的表現能力。除了抗癌以外，此藥物也有抗細菌、抗瘧疾等能力。本篇論文研究發現此藥物會抑制細胞的細胞生長(Akt)和癌幹細胞球體生成(CD133)。在腫瘤生長方面，醣基化參與細胞對蛋白質的折疊修飾，關係到蛋白質功能的表現。本篇論文也發現此藥物會有效影響N-link醣基化的能力，其中癌幹細胞指標CD133和Akt的上游基因，受體酪氨酸激酶(Receptor Tyrosine Kinase)都有發現因為抑制醣基化導致分子量下降的情況，用電腦進行分子模擬發現，此藥物會與DPAGT1這個N-link醣基化的初始酶交互作用而影響醣基化活性。海洋藥物也會和Casein kinase2的protein kinase domain交互作用，可以阻斷Casein kinase 2對PTEN磷酸化表現來達到提升PTEN的表現能力來抑制Akt的訊息傳遞路徑。在腫瘤免疫系統方面，此藥物可以藉由抑制免疫檢查點PD-L1的表現量和N-link醣基化，進而影響調控型T細胞Tregs抑制免疫系統的能力來促進免疫系統的運作。從實驗的數據顯示，此化合物對肝癌展現了強大的治療能力，可以抑制細胞生長、癌幹細胞球體形成和促進免疫系統活化來抑制肝癌細胞的生長。動物實驗抑制腫瘤大小，免疫染色中抑制細胞增生並促進細胞凋亡,腫瘤中抑制調控型T細胞。因此海洋二次代謝物可藉由N-LINK唐基來抑制癌幹細胞和免疫檢查點,除此之外可以抑制CK2-PTEN-AKT的訊息傳遞路徑來達到抑制癌細胞增生與促進細胞凋亡的效果。

Since ancient times, humans have used many terrestrial animals as a medical application, but so far there are still many diseases still cannot be cured. Due to the special creatures living in the ocean, they need to adapt to high pressure, high salt, and hypoxia habitat. Thus, it has a special chemical structure. Although human used of marine resources for drug screening time was much less than the land, but the success stories of marine drugs was much higher than the land resources, and now there are many marine drugs used in clinical medicine, I believe that the ocean resources can resolve the problems that human beings cannot solve the diseases nowadays.
Liver cancer is one of the top ten causes of cancer in Taiwan, the only liver cancer target drug sorafenib, had poor liver cancer survival rate after long-term observation, so the development of new drugs is an important project. This study is a secondary metabolite extracted from marine bacteria and assesses its ability to inhibit the growth of hepatocellular carcinoma cells.
Previous studies have found that this marine secondary metabolite in colorectal cancer for p53 mutations have a significant effect and enhance the performance of p73. In addition to anti-cancer, the drug also has anti-bacterial, anti-malarial and other capabilities. This paper investigates that this drug inhibited cell growth (Akt) and cancer stem cell spheroid formation (CD133). In tumor growth, glycosylation involved in cell folding modification of the protein, related to the performance of protein function. This paper also found that this drug would effectively affect the ability of N-link glycosylation, in which cancer stem cell markers CD133 and Akt upstream gene, receptor tyrosine kinase (Receptor Tyrosine Kinase) had been found that inhibition of glycosylation Molecular weight shifting, the use of computer simulation found that the drug would dock on DPAGT1, the N-link glycosylation initial enzyme, and affect the glycosylation activity. Moreover, marine drug could dock on Casein kinase2 protein kinase domain and block p-PTEN expression to achieve the ability to enhance the performance of PTEN to inhibit CK2-PTEN-Akt signaling pathway. In tumor immune system, this drug can promote the operation of the immune system by inhibiting immune-checkpoint PD-L1 and N-link glycosylation, thereby influencing the ability of regulatory T cell Tregs to inhibit the immune system. Animal experiments inhibited tumor sizes; immunostaining inhibited cell proliferation and promoted apoptosis and suppressed regulatory T cells. Therefore, the secondary metabolites of the ocean can inhibit the cancer stem cells and immune check point by N-linked glycosylation. In addition, suppressed CK2-PTEN-Akt signaling pathway to inhibit cell proliferation and promote cell apoptosis effect.

目錄
論文審定書 i
論文公開授權書 ii
中文摘要 iii
ABSTRACT v
INTRODUCTION 1
RESULTS 15
DISSCUSION 29
REFERENCES 70

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