本研究是藉由HSPC、膽固醇和DOTMA分子組成的微脂體(liposome)包覆碳酸氫銨(ammonium bicarbonate) 飽和溶液，擬做為一新型的癌症高溫治療系統。實驗設計是利用碳酸氫銨溶液在加熱之後會分解成為二氧化碳氣體現象，先讓癌細胞吞噬碳酸氫銨微脂體，再進行加熱步驟，提高微脂體周圍環境溫度，造成微脂體內部體積膨脹，最終導致微脂體爆破，讓癌細胞受到物理性傷害而死亡。此實驗設計可以避免癌症化學性治療方法常帶來的副作用，以及癌細胞產生抗藥性的情形。碳酸氫銨分解成為二氧化碳的溫度是在36℃~60℃間，在體溫37℃下，因為碳酸氫銨分解速度較慢，二氧化碳氣體膨脹時產生的壓力不足以造成細胞死亡。然而，由MTT assay和LDH assay實驗證明，餵食細胞1mg/ml濃度的碳酸氫銨微脂體10分鐘後，在攝氏42℃(細胞溫度耐受上限)下加熱15分鐘，會導致細胞存活率下降至約50%。如果將這些經過加熱處理、已受到物理性傷害的癌細胞，再持續培養一天後，細胞會因進入寧靜狀態(quiescent state)而使其存活率更加下降至約30%。由流式細胞儀(Flow Cytometry)分析的結果發現，這些經過物理性傷害的癌細胞中，會有40%的比例停留在G0 phase，顯示此治療方法的確能減緩癌細胞的生長。另外，由細胞對微脂體胞吞機制與其在胞器內追蹤之實驗結果，可得知在經過餵食微脂體10分鐘後，再加上15分鐘的加熱時間，微脂體會經由caveolae的傳送路徑停留在early endosome與lysosome胞器。而微脂體在lysosome胞器內的破裂，會造成溶素體酵素溶解細胞膜，對細胞造成物理性傷害。因此，此治療方式是會造成癌細胞壞死(necrosis)而不是讓癌細胞進行自發性凋亡(apoptosis)。

中文摘要 I
目錄 III
圖索引 V
表索引 VII
第一章 緒論 1
1-1 前言 1
1-2微脂體文獻回顧 2
1-3腫瘤治療方法文獻回顧 9
1-3-1.手術 9
1-3-2.化學治療 9
1-3-3.放射治療 10
1-3-4.免疫療法 10
1-3-5.高溫治療 11

第二章 範圍與研究目的 13

第三章 實驗藥品與儀器 15

第四章微脂體的基礎物性 17
4-1研究目的 17
4-2材料與方法 17
4-2-1微脂體的製備 17
4-2-2微脂體釋放量 18
4-2-3微脂體的粒徑大小和帶電量 18
4-2-4碳酸氫銨包覆率測試 19
4-2-5微脂體胞飲量測試 19
4-2-6藉由超音波影像儀測試碳酸氫銨微脂體產氣能力 19

4-3實驗結果與討論 20
4-3-1微脂體的粒徑大小 20
4-3-2微脂體釋放程度 22
4-4結論 28

第五章細胞實驗 30
5-1研究目的 30
5-2材料與方法 30
5-2-1 MTT 細胞活性染色法 30
5-2-2乳酸去氫酶活性測定 31
5-2-3 微脂體和胞器膜融合測定 31
5-2-4 碳酸氫銨微脂體副產物毒性測試 32
5-3實驗結果與討論 32
5-3-1微脂體的胞飲率 32
5-3-2最佳化測試 34
5-3-3物理性傷害對癌細胞之影響 40
5-4結論 44

第六章細胞死亡的原因探討 45
6-1研究目的 45
6-2材料與方法 45
6-2-1流式細胞儀 45
6-2-2共軛焦顯微鏡 46
6-3實驗結果與討論 47
6-3-1. 細胞死亡之分類 47
6-3-2細胞存活率 49
6-3-3細胞胞飲微脂體傳送路徑 50
6-4結論 53

第七章 Reference 54

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