臺灣博碩士論文加值系統

　　乳癌為台灣女性癌症發生率第一位，如何抑制癌細胞生長是重要的課題。本研究取二十種台灣植物之50%甲醇迴流萃物，以兩株乳癌細胞（MDA-MB-231和 MCF-7細胞株）作為靶細胞，進行體外細胞毒性檢測，並配合管柱分離，預期得到具有抑制癌細胞生長之起始天然物，結果顯示薑科植物閉鞘薑（Costus speciosus (Koenig) Smith）之根莖部抑制癌細胞生長作用較為明顯，繼而以水和乙酸乙酯作分部分析，經由Diaion HP-20管柱，以不同比例的水和甲醇沖提，分離得到5個劃分部，其中D80M和D100M具有較強的細胞毒性，且D100M具有意義的延長P-388D1之CDF1擔癌鼠存活時間。因此再利用VersaPak® Flash、Silica gel等不同分離管柱，純化D80M和D100M得到6個主要成分，分別為(25R)-neospirost-4en-3-one (1)、25-hydroxyspirost-4-en-3-one (2)、β-D-Glucopyranoside, (3β, 25R)-17-hydroxyspirost-en-3-yl 2-Ο-(6-deoxy-α-L-mannopyranosyl) (3)、Prosaponin A of dioscin (4)、dioscin (5)和gracillin (6)。其中又以Prosaponin A of dioscin、dioscin、gracillin對人類乳癌細胞具有細胞毒性，IC50分別對MDA-MB-231細胞株分別為4.88、3.43、5.37 μM，對MCF-7細胞株分別為4.6、3.39、3.36 μM，且會使乳癌細胞PARP降解，引起凋亡。另外，dioscin亦能有意義的延長P-388D1之CDF1擔癌鼠存活時間。綜合上述，閉鞘薑之薯蕷皂苷類化合物於體內、外實驗皆有活性，能引起乳癌細胞凋亡，而dioscin可作為閉鞘薑抗癌活性之指標成分或做為開發抗癌藥物的起始結構，為一個值得發展的抗癌經濟作物。

考試委員名錄
誌謝
目錄………………………………………………………………………I
圖目錄…………………………………………………………………III
表目錄…………………………………………………………………V
縮寫表…………………………………………………………………VII
中文摘要…………………………………………………………………1
英文摘要…………………………………………………………………3
緒論………………………………………………………………………5
實驗目的…………………………………………………………………26
實驗部份…………………………………………………………………27
1.材料……………………………………………………………………27
2.儀器……………………………………………………………………30
3.實驗方法………………………………………………………………31
3-1. 二十種台灣原生種植物成分萃取………………………………31
3-2. 細胞毒性篩選方法………………………………………………31
3-3. 閉鞘薑根莖部成分萃取、分離與純化…………………………34
3-4. 閉鞘薑成分之結構鑑定…………………………………………38
3-5. P-388D1誘導擔癌鼠之體內抗癌模式…………………………38
3-6. 西方點墨法分析相關蛋白的表現………………………………39
3-7.閉鞘薑萃取物指紋圖譜之分析…………………………………41
4.實驗結果……………………………………………………………42
4-1.二十種植物對乳癌細胞株之抑制活性…………………………42
4-2.閉鞘薑各部位之細胞毒性試驗…………………………………43
4-3.閉鞘薑根莖部之結構解析………………………………………46
4-4.閉鞘薑萃取物指紋圖譜之分析…………………………………51
4-5.閉鞘薑誘導之細胞凋亡…………………………………………62
4-6.閉鞘薑對P-388D1誘導擔癌鼠之體內抗癌活性試驗……………68
討論……………………………………………………………………74
參考文獻………………………………………………………………78
附錄……………………………………………………………………84

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