本研究開發脂質/中孔矽殼包覆硫化鉍之奈米藥物，搭載SN38與rapamycin兩種藥物，結合化學及放射治療應用於胰臟癌之治療。本研究利用以水熱法合成硫化鉍奈米棒，並利用中孔矽殼修飾提供拓樸異構酶抑制劑一型SN38裝載的空間，再透過脂質薄膜的包覆搭載mTOR抑制劑rapamycin並提升載體的膠體穩定性。研究中製備出的奈米載體具有均一大小的尺寸且藥物包覆率可達70%以上，亦在實驗中證實此載體具有電腦斷層(CT) 之顯影能力。細胞實驗中顯示此奈米載體可透過胞吞作用進入胰臟癌細胞UN-KC-6141中，透過細胞毒性分析確認載體所搭載之藥物可有效毒殺癌細胞，結合放射治療照射抑制細胞的增殖能力。此外，動物實驗中發現奈米載體可經由循環系統累積至腫瘤區域。結合以上成果，本研究所開發搭載雙藥物之奈米載體，其顯影能力可輔助胰臟癌之診斷，並結合化學/放射複合療法，抑制癌細胞的生長。

Our research aims to develop a lipid/ mesoporous silica coated Bi2S3 nanorod which was loaded with SN38 and rapamycin. In combination of chemotherapy and radiotherapy for pancreatic cancer treatment. In this study, we synthesized bismuth sulfide by hydrothermal method, then coated with mesoporous silica for SN38 loading. We coated lipid film to improve colloidal stability, and loaded rapamycin at the same time. The drug loaded nanoparticles showed a uniform size distribution with drug loading efficiency over 70% and contrast enhancement in CT imaging. In vitro data proved that our nanoparticles enter UN-KC-6141 cells by endocytosis, and killed the cells by the drug which were carried by the particles. In addition, in the subcutaneous pancreatic cancer model showed the nanoparticles were able to accumulate in the tumor. In conclusion, we developed a nanomedicine helpful for pancreatic cancer diagnosis by the CT contrast enhanced ability, and combined chemo and radiotherapy to inhibit cell proliferation.

摘要 i
Abstract ii
目錄 iii
表目錄 v
圖目錄 vi
一、研究動機 1
二、文獻探討 3
2.1胰臟癌 3
2.1.1胰臟癌化學治療藥物介紹 5
2.1.2胰臟癌腫瘤微環境 6
2.1.3 Enhanced Permeation and Retention Effect (EPR effect) 8
2.2奈米藥物載體傳遞系統 9
2.2.1 硫化鉍 (Bi2S3) 金屬奈米載體之特性 10
2.2.2 中孔矽殼包覆奈米載體之應用 12
2.2.3 脂質修飾奈米載體之應用 13
2.3 癌症治療策略與藥物介紹 15
2.3.1 化療藥物7-Ethyl-10-hydroxycamptothecin (SN38) 15
2.3.2 mTOR抑制劑雷帕黴素 (rapamycin) 17
2.3.3放射治療 (Radiotherapy, RT) 18
三、研究方法 21
3.1 奈米載體製備 21
3.1.1硫化鉍奈米棒 (Bismuth sulfide nanorods, Bi2S3) 合成 21
3.1.2 中孔矽殼包覆Bi2S3 (mesoporous silica coated MS-Bi2S3, MB) 合成 21
3.1.3 脂質包覆MS- Bi2S3 (lipid coated MS-Bi2S3, MB@lipid) 合成 22
3.1.4 藥物載體之製備 22
3.2 奈米載體性質探討 23
3.2.1奈米載體粒徑分布與表面電荷分析 23
3.2.2 奈米載體電腦斷層 (Computed tomography, CT) 顯影效果分析 24
3.2.3奈米載體穩定性分析 24
3.2.4 奈米藥物載體之藥物裝載量分析 24
3.2.5 奈米載體元素濃度分析 25
3.3 體外細胞實驗 25
3.3.1 細胞來源及培養環境 25
3.3.2 細胞培養液及磷酸鹽緩衝溶液配置 25
3.3.3 細胞繼代 26
3.3.4 細胞計數 27
3.3.5 細胞對奈米載體吞噬情形分析 27
3.3.6 螢光顯微鏡觀察細胞對奈米載體吞噬情形分析 28
3.3.7 細胞毒性分析 29
3.3.8 放射治療對細胞分裂影響之分析 30
3.4 動物實驗 31
3.4.1動物來源 31
3.4.2 皮下胰臟癌腫瘤建立 31
3.4.3動物體內奈米載體累積分析 31
3.5 數據統計 32
四、結果與討論 33
4.1 奈米載體性質分析 33
4.1.1 奈米載體性質鑑定 33
4.1.2 奈米載體之電腦斷層 (CT) 顯影效果 38
4.1.3 奈米載體在體外環境下之粒子穩定性 39
4.2 體外細胞實驗 40
4.2.1 細胞對奈米載體之吞噬情形 40
4.2.2 奈米載體之細胞毒性分析 43
4.2.3 放射治療對細胞分裂影響之分析 45
4.3 動物實驗 46
4.3.1 奈米載體於生物體內累積分布情形 46
五、結論 48
六、參考文獻 49

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