摘要
海洋的豐富資源為人類疾病藥物發展提供許多重要資訊。 在本研究中，從海洋生物萃取出的天然物及衍生物，評估其抑制肝癌細胞生長的能力。 結果顯示，由海綿Hyrtios sp.分離出來的化合物，展現出對肝癌細胞有好的抑制生長效果。 先前研究已發現此化合物能有效阻止乳癌腫瘤細胞的轉移作用，可以當作一種NFκB的抑制劑。 由於肝癌在台灣是主要造成癌症死亡的原因之一。 目前肝癌治療唯一標靶藥物sorafenib，對延長肝癌患者的存活率效果並不明顯，強調了開發肝癌新藥的需求。 因此本研究探討此化合物在治療肝癌上的機轉，以及透過動物模式評估其藥物開發潛力。
本論文研究發現此化合物會提高細胞週期中Pre-G0期，來達到抑制癌細胞生長並且抑制了癌細胞的侵襲及群落效應。 他可能透過抑制NFκB的訊息傳遞來影響肝癌細胞。 他也誘導肝癌細胞、腫瘤的凋亡，抑制細胞增殖指標：Ki67。 在腫瘤生長上，血管新生的功能會讓腫瘤周圍的組織變成血管以便提供腫瘤養分來達到侵襲其他組織的目的。 因此本研究也評估此化合物對血管新生的影響。 發現他有效抑制內皮細胞的生長、轉移。 在體外血管生成實驗、大鼠主動脈環實驗以及斑馬魚幼苗實驗中也都有明顯的抑制效果，我們推測他抑制血管新生是透過抑制peNOS/eNOS及VEGFR2的表現。
先前研究發現，肝癌的治療效力與癌症幹細胞有高度相關；血管新生作用與癌症幹細胞也有關連。 本研究進一步評估此化合物對癌症幹細胞的影響。 然而，他無法抑制癌症幹細胞指標(CD133/ABCG2)的表現，因此初步推斷它可能造成細胞毒殺進而抑制幹細胞球體的形成。 總而言之，此化合物對肝癌展現出強大的治療潛力、抑制了血管新生以及對抑制癌症細胞的生長。

ABSTRACT
Marine compounds constitute a bountiful resource for drug development against human diseases. A compound derived from marine sponge Hyrtios sp. (Het), has shown excellent potency in suppressing proliferation of hepatoma cells (Huh7 and N1-S1) with the lower half-maximal inhibitory concentrations (IC50). Previous studies indicated that Het inhibited invasion and metastasis of breast cancer cells and it could be used as an apoptotic inducer via inhibiting the NFκB activation. Hepatocellular carcinoma (HCC) is one of the major causes for cancer mortality in Taiwan. The current therapeutic modalities for HCC including the target therapy drug, Sorafenib, remain unsatisfactory in prolonging the survival of HCC patients, underscoring the demand for the development of novel HCC drugs. Therefore, this study was designed to investigate the mechanism of Het on HCC cells and evaluate the therapeutic potential of Het for drug development using animal model.
We found that Het potently suppressed proliferation via Pre-G0 phase block and it also inhibited invasion, colony formation ability of HCC cells. Het attenuated NFκB signaling pathway which plays an important role in HCC cells growth. Moreover, it induced HCC cells and tumor apoptosis and inhibited Ki67 (cell proliferation marker) expression. In tumor growth process, angiogenesis would make the tissue around tumor become blood vessel to nourish itself so that it could invade other tissues. As the result, we evaluate the effect of Het on angiogenesis. We found that Het showed strong potential to suppress proliferation, and migration of endothelial cells (EA. hy926 and HUVEC). Besides, the inhibition of Het in tube formation assay, rat aortic rings assay and on the vascular development in transgenic zebrafish larva Tg(kdrl:mCherryci5-fli1a:negfpy7), indicated that Het could inhibit angiogenesis. We concluded that Het inhibited angiogenesis via attenuating peNOS/eNOS and VEGFR2 expression.
Previous studies indicated that HCC therapy efficiency is associated with cancer stemness and angiogenesis. We further investigated the effect of Het on cancer stemness. However, the results of Het could not inhibit the cancer stemness marker (CD133/ABCG2). Thus, we concluded that Het might toxicity the cancer stem cell to reduce the sphere formation. From our study, Het shows a strong therapeutic potential for HCC therapy because it had the advantage of inhibiting angiogenic activity and its high toxicity to cancer cell.

目 錄
論文審定書…………………………………………………………………………………… i
論文公開授權書………………………………………………………………………….. ii
誌謝………………………………………………………………………………………………. iii
中文摘要………………………………………………………….……………………………. iv
英文摘要………………………………………………………………….……………………. v
第 一 章 Introduction……………...….……………………………………………… 1
第 二 章 Specific amis……………….………………………………………………. 8
第 三 章 Methods and materials.………………………………………………… 10
第 四 章 Results...........………………………………………………………………… 23
第 五 章 Discussion.....………………………………………………………………. 37
第 六 章 Figures and legends..……………………………………………………. 41
第 七 章 Table……......…………………………………………………………………. 77
參考文獻……………………………………………………….……………………………….. 78
附錄………………………………………………………………………………………………… 82

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