癌症為人類主要死亡原因之一，而晚期癌症往往因為腫瘤過大無法直接摘除，需要輔以化學治療，放射線治療等的治療方法，抑制腫瘤生長減小體積。而化療手段往往不能達到預期效果的原因是部分的抗癌藥物是藉由氧氣來產生自由基，並藉由自由基攻擊癌細胞來達到治療效果。但在癌細胞增殖過快導致腫瘤內部嚴重缺氧的情況下，抗癌藥物就無法產生足夠的自由基，導致癌細胞在腫瘤內部的缺氧環境下抗藥性增高，進而無法達到預期的療效。在此研究我們藉由一可植入式持續產氧系統來減緩癌細胞內部的缺氧情形，降低藥物因為缺氧而造成的治療阻力，進而提升治療效果。此產氧系統是利用褐藻酸鈉和鈣離子的交聯來包覆過氧化鈣和過氧化氫酶形成微球體，過氧化鈣與水接觸後會開始分解，過氧化氫在過氧化氫酶的催化下會大量地產生氧氣。在有氧環境及缺氧環境下進行細胞實驗，由實驗結果可以發現缺氧環境確實會降低癌細胞對藥物的敏感度，而當癌細胞在缺氧環境下，與此產氧系統共培養，抗癌藥物確實能夠產生較多自由基，進而殺死較多的癌細胞。並且進一步藉由動物模型來驗證此產氧系統是否能夠提升化學治療效果，由實驗結果我們可以發現，此產氧系統確實能夠改善缺氧環境造成的抗藥性問題，並增加抗癌藥物的毒殺效果來達到更好的治療效果。

Complete resection is the only potentially curative treatment for late stage cancer. In general, however, resection is beneficial only when there is no distant spread of disease and when all tumor can be safely removed with an adequate hepatic remnant for recovery. Thus, adjuvant therapy, such as radiotherapy and chemotherapy, has been given to shrink the tumor. Recently, hypoxia has been described as an important factor to chemotherapeutic resistance, owing to the redox state, meaning that oxygen (O2) is required to generate ROS to be maximally cytotoxic. To address the above issue, an oxygen generating system was fabricated using alginate, having calcium peroxide as the oxygen generating source, which decompose hydrogen peroxide and release oxygen when contact with water. The decomposition rate could be increased by the addition of catalase, a common enzyme found in nearly all living organisms. This novel system was performed under two controlled conditions, normoxia and hypoxia. Optimization of microspheres, cytotoxicity of material and condition of the system were carried out specifically based on the responses observed from studies using Hep3B cells as a candidate. Particles are capable of sustainably release oxygen over 24 hours while the best cross-linking time during the gelation was found to be 10 minutes. It was observed that cells maintained high viability under hypoxic condition and found that doxorubicin-induced oxyradicals play a vital role in the development of drug resistance. However, this oxygen generating system has successfully reduced the chemotherapeutic resistance which induced by hypoxia and enhanced the chemotherapeutic efficacy of doxorubicin. These results suggest that the developed oxygen generating system is a highly promising approach in maximizing the therapeutic effectiveness with minimal side effects.

目錄
致謝 I
摘要 II
ABSTRACT III
目錄 IV
圖索引 VI
公式索引 IX
第一章 緒論 1
1.1 癌症及其治療 1
1.2 腫瘤內的缺氧現象及治療抗性 2
1.3 高壓氧治療 HBO 3
1.4 過氧化氫H2O2與過氧化氫酶catalase的作用 4
1.5 研究目標 7
第二章 實驗材料及方法 10
2.1 實驗材料 10
2.2 CaO2-alginate微球的製備及表徵 10
2.2.1 CaO2-alginate微球的製備 10
2.2.2 體外缺氧環境的構建 11
2.2.3 氧氣釋放監測 12
2.2.4 H2O2 釋放及溶液pH值測定 12
2.2.5 CaO2的反應情形表徵 13
2.3 細胞實驗 13
2.3.1 材料毒性測試 13
2.3.2 CaO2-alginate 微球缺氧改善能力分析 13
2.3.3 DOX的細胞內積累及細胞抗藥性測試 14
2.3.4 關於DOX藥效增強的分析 15
2.3.5 ROS細胞內積累及細胞凋亡 15
2.4 動物實驗 16
2.4.1 建立缺氧腫瘤模型 16
2.4.2 腫瘤缺氧情形分析 16
2.4.3 關於動物實驗DOX藥效增強的分析 17
第三章 實驗結果與討論 19
3.1 CaO2-alginate微球的特徵及性質 19
3.1.1 CaO2-alginate微球的形態 19
3.1.2 氧氣釋放曲線 21
3.2 細胞實驗 23
3.2.1 材料毒性測試 23
3.2.2 產氧系統對缺氧狀態的改善 24
3.2.3 DOX的細胞內積累及治療抗性測試 26
3.2.4 產氧系統對於治療效果的增強 28
3.2.5 ROS細胞內積累及細胞凋亡 29
3.3 動物實驗 30
3.3.1 腫瘤缺氧情形改善 30
3.3.2 抗腫瘤治療效果 31
3.3.3 FDG攝取效率 33
3.3.4 腫瘤內部細胞形貌及細胞凋亡情形 34
第四章 結論 36
參考文獻 37

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