臺灣博碩士論文加值系統

近年來生活及飲食習慣的改變使得國人罹患大腸癌(colon cancer)的比例有越來越高的趨勢，而大腸癌雖然有接近90%的五年存活率，但也有很高的轉移率，使得大腸直腸癌成為世界癌症死亡的第四名。Epirubicin 為蔥環類(anthracycline)藥物，常用於治療淋巴癌、血癌、大腸癌。正常細胞內都具有一定水平的活性氧物質來調控細胞的正常生理機制，當提高或降低活性氧物質到細胞不能承受時，就可能造成細胞凋亡。沒食子酸丙酯(propyl gallate)為人工合成的化合物，可以清除自由基具有抗氧化的能力。先前研究指出沒食子酸丙酯與 Epirubicin 共同處理大腸癌細胞株 DLD-1 後，發現沒食子酸丙酯可以加強 Eepirubicin 所誘導的細胞凋亡。本篇研究希望使用不同的抗氧化劑搭配 Epirubicin 處理大腸癌細胞株，加強化療藥物的效果。在大腸癌細胞 DLD-1 和 HT-29 中，共同處理抗氧化劑沒食子酸甲酯或抗壞血酸與epirubicin的環境下都會降低細胞存活率。沒食子酸、N-乙酰半胱氨酸、榖胱甘肽共同處理epirubicin會增加 DLD-1 細胞週期 Sub G1 的比例；共同處理 Epirubicin 與抗壞血酸、N-乙酰半胱氨酸、榖胱甘肽會增加 HT-29 細胞週期 Sub G1 的比例。共同處理 Epirubicin 與N-乙酰半胱氨酸會提高 DLD-1 與 HT-29 細胞內的 RO S的產生，而共同處理 Epirubicin 與抗壞血酸則會降低 HT-29 細胞內 ROS。

In recent years, changing in living and eating habits makes people get colon cancer and have a higher and higher proportion. Although colon cancer has close to 90% of the five-year survival rates, but it also has high metastasis rates, and makes colon cancer to be the fourth in the cancer death worldwide. Epirubicin, an anthracycline, is used to treat lymphoma, leukemia, and colon cancer. In normal conditions, moderate oxidative stress in the cells regulates physiological mechanism. Elevates intracellular oxidative stress over-production or low levels may make cell apoptosis. Propyl gallate is a synthetic compound, may have antioxidant ability to remove free radical. In the previous study, propyl gallate combined with epirubicin to treat colon cancer cell line, and found that propyl gallate could enhance epirubicin-mediated apoptosis. In this study, using antioxidants combines with epirubicin treatment, enhances the effect of antitumor ability. Co-treatment with epirubicin and antioxidants propyl gallate or ascorbic acid can decrease cell viability ratio in colon cancer cell line DLD-1 and HT-29. Incombination with epirubicin and gallic acid, N-acetylcystenine or glutathione increases cell cycle subG1 ratio in DLD-1, and incombination with epirubicin and ascorbic acid, N-acetylcystenine or glutathione increases cell cycle subG1 ratio in HT-29. Co-treatment with epirubicin and N-acetylcystenine increases intracellular ROS levels in DLD-1 and HT-29, but co-treatment with ascorbic acid decreases intracellular ROS levels in HT-29.

目　　錄
中文摘要 I
Abstract II
致　　謝 III
目　　錄 IV
圖 目 錄 VIII
第一章 緒論 1
第一節 前言 1
第二節 使用藥物之簡介 2
第三節 細胞週期的調控 3
第四節 細胞凋亡的介紹 4
4.1細胞壞死 4
4.2細胞自噬 4
4.3細胞凋亡 (內因性、外因性、內質網) 5
4.4細胞凋亡相關蛋白/抗細胞凋亡相關蛋白 6
第五節 細胞內活性氧物質的簡介 8
第六節 抗氧化劑介紹 9
6.1沒食子酸(Gallic acid，GA) 10
6.2沒食子酸甲酯（Methyl gallate，MG） 10
6.3沒食子酸丙酯(Propyl gallate，PG) 10
6.4 N-乙酰半胱氨酸(N-acetyl cysteine，NAC) 11
6.5穀胱甘肽(Glutathione，GSH) 11
6.6抗壞血酸(Ascorbic acid) 11
第二章 研究目的 13
第三章 材料與方法 14
第一節 材料 14
一、細胞株 14
二、細胞培養 14
三、使用藥品 15
四、細胞存活率測試 15
五、細胞週期與細胞內活性氧及榖胱甘肽檢測 15
六、西方墨點法 16
七、抗體 17
八、軟體分析 17
第二節 方法 18
一、細胞培養 18
二、細胞存活率測試 (MTT assay) 19
三、抽取蛋白質(總蛋白質) 19
四、抽取蛋白質(核蛋白質) 19
五、蛋白質定量分析 20
六、西方墨點法 20
七、流式細胞儀 22
八、統計分析 24
第四章 結果 25
第一節 觀察 Epirubicin 濃度對大腸癌細胞 DLD-1 存活率之影響 25
第二節 觀察處理不同濃度的各種抗氧化劑對大腸癌細胞 DLD-1 存活率之影響 25
第三節 觀察 Epirubicin 與不同的抗氧化劑共同處理對大腸癌細胞 DLD-1 存活率之影響 26
第四節 觀察 Epirubicin 與不同的抗氧化劑共同處理對大腸癌細胞 HT-29 存活率之影響 28
第五節 觀察 Epirubicin 與不同的抗氧化劑共同處理對大腸癌細胞 DLD-1 與 HT-29 細胞週期的影響 29
第六節 觀察 Epirubicin 與不同的抗氧化劑共同處理對大腸癌細胞 DLD-1 與 HT-29 細胞內活性氧物質的影響 31
第五章 討論 33
第六章 圖表 41

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