前列腺癌是全世界惡性腫瘤之一，己經被證實具有腫瘤轉移的能力，會轉移侵入骨頭、膀胱、直腸和淋巴結。前列腺癌主要發現在男性成人的身上，東西方國家其致死率皆高，因此開發治療前列腺癌之化學療法為一重要的醫學研究。從篩選的398種小分子藥物中，由MTT結果顯示TCH 的衍生物明顯地對PC-3前列腺癌細胞生長抑制。此外， PC-3細胞經過TCH-1038w處理12-48小時之後進行形態學上的變化分析，我們觀察到PC-3細胞形成圓狀、細胞收縮、細胞膜皺縮等外觀的表現與細胞凋亡外觀相似。利用流式細胞儀分析我們確認了TCH-1038w會導致PC-3細胞DNA斷裂而造成sub-G1比例增加。對於觀察TCH-1038w是否造成PC-3細胞凋亡，我們以DAPI螢光染色法分析PC-3細胞死亡後之DNA完整性，發現PC-3細胞呈現DNA裂解，與流式細胞儀之結果一致。。更進一步利用西方墨點轉漬法來確認細胞凋亡相關蛋白質的表現，我們發現了procaspase-3和PARP均被切割而活化在TCH藥物處理後，顯示TCH-1038w處理後的PC-3細胞是經由細胞凋亡路徑而死亡。 以上結果我們認為TCH-1038w未來將可發展為藉誘導細胞凋亡之抗前列腺癌症之藥物。

Prostate cancer is one of the most malignant tumors in the world. It has been demonstrated that prostate cancer could metastasis to bones, bladder, rectum, and lymph nodes. It is the most common type of cancer found in adult males, and the mortality is elevated. From screening our chemical library of 398 small molecule compounds, we have found that TCH derivatives showed anti-proliferative activities using prostate cancer cell line PC-3. The results of MTT assay allow us to identify TCH-1038w as potential candidat for developing anticancer drug. Morphological investigation on PC-3 cells after TCH-1038w treatment showed that PC-3 cells rounded up and combined with cell shrinkage, abridge, membrane blebbing. Our results of flowcytometric analysis showed that TCH-1038w can cause the percentage increase of sub-G1 that indicated the DNA fragmention in TCH treated PC-3 cells. By DAPI staining , we observed that TCH-1038w can induce the DNA fragmentation in PC-3 cells. Moreover by immunoblotting analysis, we have demonstrated that procaspase 3 and PARP were cleavaged and activated after TCH treatment. These results indicsted TCH drugs can induce caspase activation, DNA fragmentation, and consequently cause apoptotic cell death. Together, TCH-1038w may serve as a potential chemotherapy candidate for treating prosate cancer in the future.

中文摘要……………………………………………………..……………...…..3
英文摘要………………………………………………….…….…………....….4
縮寫………………………………………………………………………...……5
導論………………………………………………………………………...……6
前列腺的生理結構和功能...............................................................................6
前列腺癌致病的機轉……………………………………………………...…6
前列腺癌的流行病學與診斷………………………………………...………7
前列腺癌的治療方法………………………………………………………...8
Caspase在細胞凋亡（apoptosis）所扮演的角色………………….....……10
PARP在細胞凋亡所扮演的角色……………………………..………….....15
材料和方法………………………………………………………………..…....18
人類前列腺癌細胞株PC-3和實驗藥物……………………………..…..…18
細胞生長抑制活性分析………………………………………………......…18
細胞形態學觀察……………………………………………………...…...…19
細胞週期的分析…………………………………………………………......20
螢光染色分析………………………………………………………….….…22
西方墨點轉漬（Western blotting）分析…………………………….…..…23
結果……………………………………………………………….………………....25
討論………………………………………………………………….…………...….30
Tables and Figuires………………….……………………………………..……..….33
Table 1. IC50 values of TCH drugs……………………………………..…………...34
Table 2. Effect of TCH-1038w on PC-3 cells growth…..………………..................35
Table 3. PC-3 cell cycle disruption by TCH-1038w is a dose dependant manner......36
Table 4. PC-3 cell cycle disruption by TCH-1038w is a time dependant manner…..37
Fig. 1. TCH-1038w affect on morphological change of PC-3 cells in a dose dependant manner……………………………………………………………...…….38
Fig. 2. Time- and dose-dependant inhibition of TCH-1038w in PC-3 cells………....43
Fig. 3. Dose-dependant effect of TCH-1038w on DNA fragmentation……….….....44
Fig. 4. Time-dependant effect of TCH-1038w on DNA fragmentation…..………....46
Fig. 5. Fluorescence photomicrographs of PC-3 cells with DAPI staining…….…....49
Fig. 6. Activation of caspase 3 and PARP proteins in TCH treated PC-3 cells…..…55
References…………………………………………………………………........…...57

行政院衛生署衛生統計資料網歷年男性癌症死亡原因，95年度，表 14。
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