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系統識別號 U0026-0812200910213246
論文名稱(中文) Azatyrosine衍生物HPW 99系列抗癌活性之研究
論文名稱(英文) An Investigation on the Anticancer Activities of Azatyrosine Anlogues-the HPW 99 series
校院名稱 成功大學
系所名稱(中) 藥理學研究所
系所名稱(英) Department of Pharmacology
學年度 90
學期 2
出版年 91
研究生(中文) 王靜敏
研究生(英文) Ching-Min Wang
學號 s2689407
學位類別 碩士
語文別 中文
論文頁數 69頁
口試委員 指導教授-李益謙
口試委員-王惠珀
口試委員-賴明德
中文關鍵字 細胞移行  轉移  血管新生 
英文關鍵字 aztyrosine  angiogenesis. metastasis  CAM assay 
學科別分類
中文摘要 中文摘要

Azatyrosine [L-β-(5-hydroxy-2-pyridyl)-alanine]為1989年由Yoshiaki等人從Streptomyces chibanesis 中所分離出來,具有抗癌作用的抗生素。在1992年,Izawa等人發現azatyrosine可以減少小鼠因DMBA誘導,而產生的乳突狀腫瘤的面積和大小。由於azatyrosine需要在高劑量下才具有抗癌活性,因此,台大及長庚大學的王惠珀教授以azatyrosine為leader compound,將azatyrosine的結構加以修飾合成了一序列化合物 ( HPW 98、 99序列)。本實驗室過去曾發現,HPW 98序列在抗癌活性比azatyrosine強,但不會像azatyrosine一樣在合成蛋白質時與tyrosine競爭。在本項實驗中我更進一步針對HPW 99序列中的HPW 99-5,探討其相關的抗癌活性。結果顯示,HPW 99序列藥物在一些人類癌細胞中具有良好的細胞毒性,且此種毒殺作用是作用於細胞週期中,我們也發現,HPW 99-5會使細胞停留在G2/M期,並會造成細胞走向凋亡。但HPW 99-5與我們所選用的抗癌藥物進行併用療法 (combination therapy),似乎對大腸直腸癌細胞SW480與前列腺癌細胞PC-3,不具有累加作用。在抗血管新生的活性上,發現HPW 99-5除了可以抑制內皮細胞BAEC (bovine aortic endothelial cell)的增生,也影響細胞移行 (cell migration)之能力。使用雞胚胎卵黃囊試驗法 (chicken chorioallantoic membrane ,CAM assay),來觀察藥物在活體中的活性。但發現藥物必須在相當高的濃度下,才可能具有抑制血管新生的作用,這可能是由於打入的藥物會被蛋中液體稀釋所導致的結果。而由於溶解度的問題,無法避免掉DMSO溶劑。對胚胎所造成的毒性。細胞移行實驗 (cell migration assay),證明HPW 99-5會抑制prostate cell (PC-3 cells)的轉移能力。在動物實驗方面,發現將NIH 3T3/7-4 cells (ras overexpression)以皮下打入Balb/c mice後,若在24小時後立即投予HPW 99-5,可以防止癌細胞的生長;若在一星期後在投予藥物,也可以抑制癌細胞繼續增生。最後使用小鼠路以士肺癌轉移模式,測試藥物是否在體內具有抑制腫瘤轉移的能力。初步發現藥物對於原位癌細胞的生長抑制情形很好,但當將原位癌細胞切除後,HPW 99-5似乎無法抑制轉移後的癌細胞,在肺臟中的生長。
英文摘要 bstract

Azatyrosine [L- β-(5-hydroxypyrid-2-yl)-alanine], an antibiotic agent isolated from Streptomyces chibanesis, is tyrosine analogue in which a pyridine group replaces the benzene group. The chemical has been reported to inhibit 7, 12-dimethyl-benz [a]-anthracence-induced carcinogenesis in mice. However, to exert its cytotoxicity, azatyrosine should be used at a fairly high concentration. The HPW 99- series are synthetic analogues of azatyrosine designed to be more lipophic for easier entry into the cells. One analogue, HPW 99-5, exhibits IC50 values lower than that of azatyrosine when used to against some cancer cell lines. The drug owes its cytotoxicity to interference of cell cycle progression. The result of propidium iodide staining indicates that the cells upon drug treatment arrest in the G2/M phase and exhibit apoptosis. The analogue dose not exhibit additive activity when combined with some anticancer drugs on the human colon cancer cell line (SW480) and human prostate cancer cell line (PC-3). We also find out that the analogue interferes the proliferation and migration of endothelial cells. In order to know if the analogue processes antiangiogenic activities, we also use the chorio- allontoic membrane assay. However, to exert its antiangiogenic activity the analogue has to be used at a fairly high concentration. The analogue also interferes the migrating capacity of human prostate cancer cells. The anticancer activity of HPW 99-5 in the established animal model is as the following. The analogue inhibits the growth of the tumor when Balb/c mice were subcutaneously injected with ras-transformed NIH3T3 cells for 24 hours later the analogue administered, the inhibition is irreversible. If the drug is administered 1week after tumor cells injected, the analogue still inhibits the tumor growth. We used mouse Lewis lung carcinoma as the metastasis animal model. The results show that the drug can inhibit the growth of the primary tumor growth, but can not inhibit the metastases growth in the lung.
論文目次 考試合格證明
誌謝
目錄 I
圖目錄 III
表目錄 V
中文摘要 VI
英文摘要 VIII
縮寫表 X
第一章 研究背景 1
第二章 研究目的 9
第三章 實驗材料 11
第一節 細胞株 11
第二節 試劑 11
第四章 實驗方法 13
第一節 細胞培養 13
第二節 離體細胞毒性 15
第三節 非增生細胞藥物毒性測試 17
第四節 合併藥物細胞毒性 19
第五節 遺傳毒性測試 21
第六節 體外癌細胞移行測試 23
第七節 內皮細胞增生試驗 24
第八節 內皮細胞移行試驗 25
第九節 雞胚胎絨毛膜試驗 27
第十節 細胞凋亡之分析 28
第十一節活體實驗 29
第五章 實驗結果 31
第六章 討論 39
參考文獻 44
圖表 48
作者簡歷 69
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