肝細胞癌是全世界最常見的惡性腫瘤第五名，同時在癌症死亡率排名第三。蕾莎瓦(Sorafenib)是第一個也是現在唯一被FDA在2007年核准用來治療晚期肝細胞癌的藥物，所以篩選出新穎藥物用於治療肝癌是非常重要且迫切的。先前實驗室已經建立斑馬魚肝癌模型可在11個月產生肝癌，本論文的第一部分為建立一個較高發生率且更早誘發肝癌的模型。藉由過度餵食的方法，我們發現HBx,src(p53-)三重轉殖基因的母魚在過度餵食下可在五個月產生肝癌。這個模式將用於之後篩選抗肝癌藥物之用。
本論文的第二部分為利用血管螢光魚(fli1：EGFP)胚胎篩選來自生技與藥物研究所提供的小分子藥物庫找出具抑制血管新生作用的小分子藥物。從LIB1F和LIB1O這兩個有抑制血管新生作用的核心結構之560種衍生物，我們發現共69種衍生物具有抑制血管新生的作用。從中挑出六種不會造成胚胎發育缺陷的小分子藥物，發現LIB1O0078和LIB1O0144具有極低的半抑制濃度(IC50)，接著測試這兩種藥物在異種移植模式抗癌細胞增生及轉移的能力，及在轉殖基因斑馬魚治療肝癌的能力。我們也測試這兩種藥物的肝毒性，以及對胚胎毒性。
我們發現LIB1O0078具有較佳的抗癌細胞增生的能力，而LIB1O0144抗癌細胞轉移的能力較好。接著我們利用HBx,src(p53-)三重轉殖基因的母魚過度餵食8週在五個月產生肝癌之動物模式，眼窩注射藥物4週，發現LIB1O0078具有抗肝癌的能力，LIB1O0144抗肝癌效果不如LIB1O0078。毒性測試方面，LIB1O0078和LIB1O0144比Sorafenib造成更低的肝毒性。胚胎毒性方面，LIB1O0078和Sorafenib造成的胚胎毒性跟非常類似，且比LIB1O0144具更低毒性。從我的論文研究中，我們發現了兩種新穎小分子對肝癌生成具有療效，且比美國FDA批准的抗肝癌的藥物（sorafenib）更安全。因此，我們的結果證明，斑馬魚平台的確是發現抗癌新藥物的一種優異模型。

Hepatocellular carcinoma (HCC) ranks as the fifth commonest of malignant tumors worldwide and the third leading cause of cancer-related death. Sorafenib is the first and only FDA-approved multi-kinase inhibitor for the treatment of advanced HCC in 2007, so screening the new drugs for treating HCC is important and urgent. Previously we had established zebrafish HCC models which developed HCC at 11 months. The first part of my thesis was to develop a HCC model which has higher incident and earlier onset. By using diet-induced obesity model, we found HBx and src in the background of p53 mutant triple transgenic female fish could develop HCC at 5 months of stage. This animal model will be used for screening drugs of anti-HCC.
The second part of my thesis was using fli1:EGFP transgenic zebrafish embryos to screen the 560 derivatives of two LIB1F and LIB1O core structures which exhibited anti-angiogenesis effect, and discovered 69 derivatives exhibited anti-angiogenesis effect. Six compounds which did not result in embryo development defect was used for titration to determine their IC50. Two compounds (LIB1O0078 and LIB1O0144) were found having the lowest IC50, and were continued to test their therapeutic effect against liver cancer using transgenic fish, and anti-proliferation and anti-migration ability using xenotransplantation model and their toxicity.
In xenotransplantation assay, LIB1O0078 exhibited better anti-proliferation ability and LIB1O0144 with better anti-migration ability. Using the 5 month old HBx and src in the background of p53 mutant triple transgenic female fish, orbital injection of drugs into blood for 4 weeks, we discovered LIB1O0078 could reverse HCC into normal. The therapeutic effect of LIB1O0144 was lesser than LIB1O0078. In terms of toxicity, LIB1O0078 and LIB1O0144 had lower hepatoxicity than sorafenib. In the survival test, LIB1O0078 and sorafenib had similar and less toxicity than LIB1O0144. From my thesis project, we discovered two novel small molecules which had therapeutic effect on HCC formation and were safer than FDA approved anti-liver cancer drug (sorafenib). Therefore, our data proved that the zebrafish platform was an excellent model for identification of new anti-cancer drugs.

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