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研究生: 王國全
Wang, Kuo-Chang
論文名稱: Canavalia galadiata地方品種抗癌活性之研究
An Investigation on the General Anticancer Activities of a local variety of Canavailia galadiata
指導教授: 李益謙
Li, I-Cjian
學位類別: 碩士
Master
系所名稱: 醫學院 - 藥理學研究所
Department of Pharmacology
論文出版年: 2003
畢業學年度: 91
語文別: 中文
論文頁數: 102
中文關鍵詞: 白鳳豆
外文關鍵詞: Canavailia galadiata
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  • 我們實驗室自坊間宣稱具有抗癌療效的豆科植物中,收集到地方品種Canavailia galadiata種子,利用不同的萃取方法,探討其粗萃取液是否具有抗癌活性。再從具有抗癌活性的部分進行純化分離,找出具有抗癌活性的級分(fraction)或指標成分,並探討其活體生物毒性及抗癌活性的機制。初步篩選到一個級分 (fraction),命名為IC-1。IC-1在一些人類癌細胞株中,具有良好的毒殺作用,細胞週期分析 (cell cycle analysis) 顯示IC-1對於癌細胞株所產生的細胞毒殺作用,可能是藉由細胞凋亡 (apoptosis) 而來的。使用IC-1和一些抗癌藥(例如:Doxorubucin,5-Fu,VP-16) 進行合併療法 (combination therapy),得知IC-1具有增強化療藥物的作用。在抑制血管新生的活性上,我們發現IC-1確實能影響到內皮細胞BAEC之移行 (cell migration) 的能力。我們也使用雞胚胎卵黃囊試驗法(chicken chorioallantonic membrane,CAM assay),來觀察IC-1在活體中抑制血管新生的情形,結果發現IC-1能有效的抑制血管新生。癌細胞移行實驗 ( cell migration assay) 證實IC-1能有效抑制PC-3的移行能力。在動物實驗方面,發現將NIH3T3/7-4 cells 以腹腔注射方式打入Balb/c mice 後,在24hr後立即給予IC-1,能有效的延長存活天數。另一模式為小鼠路易士肺癌轉移模式,目的是測試藥物在體內是否具有抑制腫瘤轉移的能力,其結果為IC-1無論對於原位腫瘤或轉移的腫瘤均具有良好的抑制作用。

    An ammonium sulfate fractioned and water soluble crude extract from a local variety of Canavailia galadiata was tested for its general anticancer activity. In vitro tests showed that the extract is toxic to some cancer cell lines in a dose-dependent manner; the toxicities, however, disappeared when the extract was heated, suggesting that the toxic principle is a protein. Cell flow cytometry study suggested that cancer cell death is due to apoptosis. In drug combination studies the extract enhanced the cytotoxicities elicited by some currently commonly used cancer chemo-therapeutics agents such as 5-FU, VP-16 or doxorubicin. The extract inhibited the migration of cancer cell (PC 3 prostate) as well as bovine aortic epithelial cell in a Boyden chamber assay. In a semi in vivo chicken chorioallantonic membrane (CAM) assay the extract inhibits blood vessel formation, suggesting that the extract possesses some anti-angiogenesis activities. Furthermore, the extract when injected intraperitoneally into testing animals was found to be able to (1) prolong the life of Balb/C mice bearing intraperitoneally injected NIH3T3/ras cancer cells; (2) reduce the tumor size when NIH3T3/ras cells was transplanted subcutaneously into Balb/C mice; (3) inhibits the growth of primary tumors in the skin and the metastatic tumors at the lung when Lewis lung carcinoma cells were transplanted subcutaneously into C57BL/6 mice.

    考試合格證明 2 誌謝 3 目錄 4 圖目錄 6 表目錄 8 中文摘要 9 英文摘要 10 縮寫表 11 第一章 研究背景 12 第二章 研究目的 23 第三章 實驗材料 25 第一節 細胞株 25 第二節 試劑 25 第四章 實驗方法 27 第一節 植物萃取 27 第二節 植物蛋白質的定量 28 第三節 植物蛋白質的純化 29 第四節 細胞培養 32 第五節 細胞計數 34 第六節 離體細胞毒性 34 第七節 合併藥物細胞毒性 37 第八節 非增生細胞藥物毒性測試 38 第九節 細胞凋亡之分析 40 第十節 體外癌細胞行試驗 41 第十一節內皮細胞移行試驗 43 第十二節雞胚胎絨毛膜試驗 45 第十三節活體動物實驗 46 第五章 實驗結果 48 第六章 討論 57 參考文獻 62 圖表 70 作者簡歷 102

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