中文摘要 Project 1：
目前在臨床上艾黴素 (doxorubicin) 是廣泛用於治療癌症的化療藥物，而長期使用艾黴素治療癌症所引起的抗藥性一直是治療癌症的障礙。近期有研究指出，化療藥物所產生的ROS (reactive oxygen species) 會促進抗藥性癌細胞的生長，然而，氧化還原的修飾和抗藥性癌細胞中那些目標蛋白質作用尚未釐清。我們藉由半胱胺酸 (cysteine) 螢光標定蛋白質樣品，進行氧化還原二維差異電泳 (Redox-DIGE) 並結合基質輔助雷射脫附游離法 (MALDI-TOF MS) 分析影響抗藥性子宮肌瘤癌細胞中氧化還原的調控蛋白。在蛋白質體學技術研究指出有33個蛋白質點其巰基團 (thiol group) 有顯著的變化，這些蛋白質參與了細胞骨架調節、訊息傳遞、氧化還原調節、糖解作用和細胞週期調節。以目前研究結果指出，氧化還原二維差異電泳提供了一個快速的方法來研究艾黴素對子宮肌瘤癌細胞的分子機制以及成為診斷治療的應用。

中文摘要 Project 2：
粒線體在細胞中是一個很重要的胞器，它負責細胞的多種功能，包括能量代謝調節、細胞生長與分裂、自由基的產生並參與細胞凋亡路徑。曾有研究指出，使用化療藥物治療的癌細胞時，會導致粒線體的缺陷和氧化磷酸化及能量產生等機能失調。在我們的研究中，是從細胞中萃取粒線體以增加粒線體蛋白質的含量，再將粒線體蛋白質樣品進行二維差異電泳 (2D-DIGE) 並結合基質輔助雷射脫附游離法 (MALDI-TOF MS) 分析在不同抗藥性的子宮肌瘤細胞之間有差異表現的粒線體蛋白質。在蛋白質體學技術研究中我們鑑定出有101個蛋白質點有顯著性變化，其中有39個蛋白質點是屬於粒線體的部分。我們進一步使用RNA干擾技術，研究乙醯輔酶A乙醯轉移酶 (ACAT1) 和蘋果酸脫氫酶2 (MDH2) 這兩個粒線體蛋白質。在我們的研究結果發現，當細胞knockdown掉這兩個粒線體蛋白質後，其細胞存活率下降，而且增加細胞凋亡的現象。由此實驗結果顯示，在未來可望在處理抗藥性子宮肌瘤癌時，ACAT1和MDH2能成為未來治療和診斷的指標。

Abstract Project 1 :
Doxorubicin is a chemotherapy used in a wide range of cancer therapies in clinical, and doxorubicin-induced drug resistance following long-time drug exposure is one of the most serious obstacles of chemotherapy. Recent studies have showed chemotherapy will produce the generation of ROS (reactive oxygen species) and then promote drug resistant cancer cells grow. However, the redox-modifications of proteins and cellular targets central to resistant cancers are largly unknown. We used the cysteine labeling protein samples by means of two-dimensional differential gel electrophoresis (2D-DIGE) and MALDI-TOF mass spectrometry (MALDI-TOF MS) to analyze the effects of doxorubicin resistance on redox-regulated proteins in uterine cancer cells. Proteomics research studies demonstrated 33 protein spots have significantly changed in thiol group reactivity. These proteins involve in cytoskeleton regulation, signal transduction, redox-regulation, glycolysis and cell-cycle regulation. In the present results, redox 2D-DIGE provides a quickly way to study the molecular mechanism and possible diagnostic or therapeutic applications in the doxorubicin-induced drug resistant uterine cancer cells.

Abstract Project 2 :
Mitochondria are key organelles, they are responsible for several cellular functions including energy metabolism regulation, cell growth, division, free radical production and participate cell apoptosis previous studies showed chemotherapy for cancer cells may lead to mitochondrial defects, oxidative phosphorylation and energy production disorders. In our study, mitochondria were isolated and mitochondrial proteins were enriched and obtained. By means of two-dimensional differential gel electrophoresis (2D-DIGE) and matrix-assistant laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS), 101protein spots were significantly changed, including 39 protein spots belongs to mitochondria. We further used RNA interference to analyze acetyl-coenzyme A acetyltransferase 1 (ACAT1) and malate dehydrogenase 2 (MDH2). Our data indicated cell viability is decreased and cell apoptosis is increased when these two mitochondrial proteins were knockdown. Experimental results show ACAT1 and MDH2 can become therapeutic candidates and diagnostic markers for the treatment of drug-resistance uterine cancer in the future.

Project 1 :
中文摘要...2
Abstract...3
壹、文獻探討...4
貳、材料與方法...34
參、實驗結果...47
肆、討論...61
伍、結論...63
Project 2 :...64
中文摘要...65
Abstract...66
壹、文獻探討...67
貳、材料與方法...75
參、實驗結果...94
肆、討論...122
伍、結論...125
參考文獻...126
附 錄...131

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