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研究生: 許琇雯
Hsiu-Wen Hsu
論文名稱: 探討薑黃素在人類大腸癌HT-29細胞之抗癌作用
Identify the effect of curcumin on colon cancer HT-29 cells.
指導教授: 蘇純立
Su, Chun-Li
學位類別: 碩士
Master
系所名稱: 人類發展與家庭學系
Department of Human Development and Family Studies
論文出版年: 2014
畢業學年度: 103
語文別: 中文
論文頁數: 91
中文關鍵詞: 大腸癌薑黃素Aurora-ANF-kB細胞凋亡細胞自噬BAF
英文關鍵詞: human colonrectal cancer, curcumin, Aurora-A, NF-kB, apoptosis, autophagy, BAF
論文種類: 學術論文
相關次數: 點閱:186下載:9
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  • 台灣衛生福利部公告,大腸癌為102年台灣癌症十大死因第三位。癌症發展過程之生物機轉複雜,現行療法無法根除癌細胞且有副作用、復發率高,因此尋找有效替代療法是非常重要的。薑黃素(Curcumin)具有低細胞毒性、多目標阻斷路徑、抑制癌症發展的特性。文獻指出,細胞凋亡、細胞自噬、發炎因子NF-B和致癌基因Aurora-A kinase之間具有相關性。本研究在於探討薑黃素對HT-29細胞週期的影響、細胞凋亡與細胞自噬的現象及NF-B與Aurora-A kinase相關蛋白的影響。結果顯示,薑黃素增加促凋亡蛋白caspase 3、細胞自噬蛋白LC3-II表現量,代表HT-29啟動細胞凋亡與細胞自噬,抑制細胞生長。NF-B相關蛋白實驗顯示細胞中IB、phospho-IB及細胞核中NF-B蛋白量顯著下降。流式細胞儀分析觀察到薑黃素增加酸性囊泡比例,使用BAF抑制細胞自噬,降低薑黃素引起酸性囊泡比例的增加。整體而言,薑黃素影響HT-29細胞週期、引發細胞凋亡及細胞自噬、抑制HT-29細胞生長,並抑制NF-B路徑及Aurora-A相關蛋白的表現;BAF的使用可降低薑黃素引起的細胞自噬。

    According to the report from the Ministry of Health and Welfare, Taiwan, 2013, human colorectal cancer is the third-leading cause of cancer death in Taiwan. Due to the complexity and high recurrent rate of cancer, as well as side effects of cancer treatment, improvement in chemotherapeutic regimens is urgently needed. Curcumin, safe in human, regulates multiple pathways to inhibit tumor progression, such as apoptosis, autophagy, inflammatory factor NF-B and oncogene Aurora-A. This study was proposed to investigate the effect of curcumin on cell cycle, apoptosis, autophagy, and protein expression and activity of NF-B and Aurora-A using human colorectal cancer HT-29 cells. The results showed that curcumin increased the expression of pro-apoptotic protein caspase 3 and autophagy protein LC3-II, suggesting that curcumin induced apoptosis and autophagy. Significant decline of IB and phospho-IBin whole cell lysates and NF-B in the nuclei were observed. Flow cytometric analysis found that the proportion of acidic vesicles increased in curcumin-treated HT-29 cells. Administration of BAF reduced curcumin-induced increment of the proportion of acidic vesicles. Overall, curcumin changed cell cycle, triggered apoptosis and autophagy, inhibited the growth of HT-29 cells, and inhibited expression of NF-B and Aurora-A. The use of BAF reduced curcumin-induced autophagy.

    第一章 緒論 1 第一節 大腸癌(Colon cancer) 1 一、 大腸癌流行病學調查 1 二、 大腸癌目前的治療 1 第二節 薑黃素(Curcumin) 4 一、 薑黃素的背景以及結構 4 二、 薑黃素生物特性 4 三、 薑黃素與大腸癌 5 第三節 細胞死亡 7 一、 細胞凋亡(Apoptosis) 7 二、 細胞自噬(Autophagy) 8 三、 細胞凋亡與細胞自噬相關性 10 第四節 Aurora-A kinase 12 一、 Aurora-A kinase背景 12 二、 Aurora-A kinase參與有絲分裂的過程 12 三、 Aurora-A kinase與癌症 13 第五節 NF-kB 14 第六節 Aurora-A kinase和NF-B相關研究 15 第二章 研究目的 16 第三章 材料與方法 18 第一節 實驗儀器與藥品耗材 18 一、 儀器與耗材 18 二、 抗體 24 三、 實驗藥品與試劑 25 第二節 分析軟體 30 第三節 實驗方法 31 一、 細胞株解凍、繼代及培養以及冷凍 31 二、 細胞計數 33 三、 藥物配置 34 四、 細胞存活分析 34 五、 細胞蛋白製備 35 六、 西方墨點法 39 七、 細胞週期分析 45 八、 細胞自噬比例分析 47 九、 統計分析方法 48 第四章 結果 49 第一節 薑黃素對大腸癌HT-29細胞生長抑制情形 49 第二節 薑黃素對於大腸癌HT-29細胞週期的影響 53 第三節 薑黃素引發細胞凋亡抑制大腸癌HT-29細胞生長 56 第四節 薑黃素影響大腸癌HT-29細胞中NF-kB的表現 59 第五節 薑黃素對大腸癌HT-29細胞中Aurora-A kinase的影響 63 第六節 薑黃素造成大腸癌HT-29細胞引發細胞自噬情形 67 第五章 討論 72 第六章 結論 77 第七章 參考文獻 78 第八章 附錄 85

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