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研究生:曾姿斐
研究生(外文):Tzi-Fei Tzeng
論文名稱:KotomolideA透過ROS/JNK的分子訊息傳遞路徑誘發人類乳癌細胞進行細胞凋亡
論文名稱(外文):Involvement of reactive oxygen species/c-Jun NH2-terminal kinase pathway in Kotomolide A induces apoptosis in human breast cancer cells
指導教授:林俊清林俊清引用關係
指導教授(外文):Chun-Ching Lin
學位類別:碩士
校院名稱:高雄醫學大學
系所名稱:天然藥物研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:80
中文關鍵詞:細胞凋亡反應性活性氧JNK乳癌細胞
外文關鍵詞:Kotomolide AapoptosisROSJNKbreast cancer
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Kotomolide A (KTA) 為Cinnamomum kotoense (Lauraceae)中所分離出之butanolide類化合物,本研究首次發現對乳癌細胞株 MCF-7 和 MDA-MB-231 具有抗癌活性,並深入探討其作用機轉 Kotomolide A (KTA)對乳癌細胞株 MCF-7 和 MDA-MB-231細胞具有抗增生活性,並可有效抑制腫瘤細胞的增生,且高度選擇性不會對正常細胞產生毒性。KTA可以藉由停止細胞週期於G2/M期並誘導細胞凋亡而有效地抑制MCF-7 和 MDA-MB-231細胞生長。KTA會藉由增加p21/WAF1的表現外,也會抑制cyclin A、cyclin B1、cdc2 和 cdc25C的表現並同時增加cdc2和cdc25C的磷酸化作用,由此,得知KTA會促使MCF-7 和 MDA-MB-231細胞週期停滯於G2/M期。另一方面,KTA會透過DR5(death receptor)和粒腺體細胞凋亡路徑進行細胞凋亡作用,而不是透過Fas receptor的活化,以KTA處理MCF-7 和 MDA-MB-231細胞,會導致細胞內Glutathione的減少與ROS的增加,並且隨後JNK的活化及產生細胞凋亡。對於ROS方面,利用兩種抗氧化劑NAC 和抗氧化酵素 catalase 作用下,可明顯抑制JNK磷酸化及DR5所誘導的細胞凋亡。另外,採用siRNA技術來降低JNK表現,同樣也可抑制KTA所調控Bim剪切及阻斷DR5上游調控所促使的細胞凋亡,而確認KTA會透過JNK活化而啟動細胞凋亡。最後,利用nude mice的實驗模式也確認KTA在in vivo條件下具有50%腫瘤生長抑制率,仍藉由活化JNK而啟動細胞凋亡的機制而能有效地抑制腫瘤細胞的生長。經由本實驗結果證實,Kotomolide A (KTA)會透過誘導ROS的生成和JNK活性,而促使人類乳癌細胞進行細胞凋亡。
The anticancer effects of kotomolide A (KTA), a new butanolide constituent isolated from the leaves of Cinnamomum kotoense (Lauraceae), on the two human breast cancer cell lines MCF-7 and MDA-MB-231, were first investigated in our study. KTA exhibited selectively antiproliferative effects in cancer cell lines without showing any toxicity in normal mammary epithelial cells. Treatment of cancer cells with KTA to trigger G2/M phase arrest was associated with increased p21/WAF1 levels and reduced amounts of cyclin A, cyclin B1, cdc2 and cdc25C. KTA induced cancer cell death treatment by triggering mitochondrial and death receptor 5 (DR5) apoptotic pathways, but did not act on the Fas receptor. Exposure of MCF-7 and MDA-MB-231 cells to KTA resulted in cellular glutathione reduction and ROS generation, accompanied by JNK activation and apoptosis. Both antioxidants, NAC and catalase, significantly decreased apoptosis by inhibiting the phosphorylation of JNK and subsequently triggering DR5 cell death pathways. The reduction of JNK expression by siRNA decreased KTA-mediated Bim cleavage, DR5 upregulation and apoptosis. Furthermore, daily KTA i.p. injections in nude mice with MDA-MB-231 s.c. tumors resulted in a 50% decrease of mean tumor volume, compared with vehicle-treated controls. Taken together, the data show that cell death of breast cancer cells in response to KTA is dependent upon ROS generation and JNK activation, triggering intrinsic and extrinsic apoptotic pathways. The ROS/JNK pathway could be a useful target for novel approaches in breast cancer chemotherapy.
目錄 Ⅰ
中文摘要 Ⅱ
英文摘要 Ⅳ
第一章、序論 1
第二章、材料與方法 18
第三章、結果 34
第四章、討論 44
第五章、參考文獻 50
第六章、圖表 61
第七章、附圖 75
第八章、學術發表 76
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