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系統識別號 U0026-0812200915352387
論文名稱(中文) 刀豆蛋白A複合的金奈米粒子對於小鼠肺癌治療之研究
論文名稱(英文) Therapeutic Effects of Concanavalin A-Conjugated Gold Nanoparticles on Lung Cancer in Mice
校院名稱 成功大學
系所名稱(中) 生物化學暨分子生物學研究所
系所名稱(英) of Biochemistry and Molecular Biology
學年度 97
學期 2
出版年 98
研究生(中文) 李國煌
研究生(英文) Kuo-Huang Li
學號 s1694108
學位類別 碩士
語文別 中文
論文頁數 55頁
口試委員 指導教授-吳昭良
口試委員-蕭璦莉
口試委員-陳毓宏
口試委員-謝嘉興
口試委員-彭國証
中文關鍵字 細胞自噬  刀豆蛋白  金奈米  膠囊效應 
英文關鍵字 Capsule Effect  Gold Nanoparticle  Autophagy  ConA 
學科別分類
中文摘要 對於專一性的標靶治療,金奈米載體提供了一個全新的平台。金奈米粒子利用特有的物理與化學的特性在藥物運送上扮演重要的角色。複合上生物性的抗原或抗體、光學感應器、疾病及癌症治療的藥物的金奈米複合物被廣泛的應用在生物體上。
ConA,一種從Jack bean 中分離出的醣類結合蛋白。在正常生理pH和溫度下,ConA靠著氫鍵和六個分子間的鹽橋形成四聚體的形式,並且對於α-D-glucose, α-D-mannose有極高專一性的親和力,可以藉由這個特性結合到細胞表面上具有此類糖蛋白的區域。另一方面,ConA在In Vitro和In Vivo上有許多生物活性,包含刺激免疫細胞增生(Mitogenesis) 和細胞毒性(Cytotoxicity)。
在我的研究裡,我製備出ConA 複合的金奈米粒子“ ConA–AuNP ”複合物,並且利用癌細胞在In Vitro 及 In Vivo上測試它的治療效果。根據我們的實驗結果發現,在由LL2細胞所引起的小鼠路易斯型肺癌上ConA–AuNP複合物相較相同劑量下的 Free ConA無論在In Vitro 以及In Vivo都有更好的治療效果。
因此,我們的認為ConA–AuNP複合物相較於Free ConA在癌症的治療上可能有更好的治療效果。
英文摘要 This study provides a novel platform for Gold nanoparticles (AuNP) as agents of target-specific delivery therapy. AuNP exploit their unique chemical and physical properties for transporting and loading the pharmaceutics. AuNP conjucated with biological ligands or antibody, optical sensing, and therapeutic drug for various diseases or in cancer is popular used in biology.
Concanavalin A (ConA) is a lectin isolated from jack bean. At physiological pH value and temperature, the tetramer predominates, which is stabilized by hydrogen bonds and six inter-dimer salt-bridges. Because ConA has high affinity to α-D-glucose and α-D-mannose, it can bind to the carbohydrate moiety of glycoconjuates on the cellular membranes. On the other hand, ConA has been associated with a variety of biological effects including mitogenesis and cytotoxicity in vitro and in vivo.
In this study, I developed a formulation of ConA bound to the gold nanoparticle (ConA–AuNP) complex and examined its therapeutic effects on cancer cells in vitro and in vivo. ConA–AuNP complex showed higher cytotoxic effects on mouse lewis lung carcinoma cells (LL2) compared with an equal dose of Free ConA.
These results suggest that ConA–AuNP complex may be more effective than free ConA for cancer treatment.
論文目次 考試合格證明
中文摘要
英文摘要
誌謝
總目錄
圖目錄
第一章 緒論
(一)、金奈米粒子(AuNP)
(二)、刀豆蛋白(Concanavalin A)
(三)、肺癌(Lung Cancer)
(四)、細胞自噬(Autophagy)
第二章 研究目的
第三章 材料與方法
(一)、材料
1、金奈米粒子
2、刀豆蛋白
3、細胞株
4、實驗動物
(二)、方法
1、ConA-AuNP複合物的製備
2、細胞生長分析
3、表面電位分析
4、微量蛋白質濃度分析
5、ConA-AuNP 奈米複合物穩定度測試
6、西方點墨法
7、TUNNEL分析
8、LDH 分析
9、LL2細胞攝入Free ConA與ConA-AuNP複合物分析實驗
10、動物實驗
11、脾臟細胞培養
12、統計分析
第四章 結果
(一)、實驗前觀察,ConA對LL2細胞生長的抑制上大於NIH3T3細胞
(二)、製備ConA複合的金奈米粒子 (ConA-AuNP)
(三)、在抑制LL2細胞生長上,ConA-AuNP 組相較其他Control組大
(四)、給予ConA-AuNP 12~24小時LL2細胞發生了Autophagy,並且大部分的細胞在24小時後逃不過Autophagy而導致細胞死亡
(五)、以ConA-AuNP的形式,可加速細胞攝入ConA的時間並且增加單位時間內細胞攝取ConA的量
(六)、In Vivo方面,ConA-AuNP也有更好的治療效果
(七)、初步排除免疫細胞參與在CL57B/6老鼠胸腔原位癌模式的治療過程
第五章 結論
第六章 討論
參考文獻
圖表
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