臺灣博碩士論文加值系統

膀胱癌在最近十年內一直是國人男性十大癌症之一(台灣男人中第七常見的癌症，美國男人中是第四常見的癌症)，也是台灣地區國人最常見的泌尿系統癌症。膀胱癌泛指各種出自膀胱的惡性腫瘤，也就是有異常細胞大量增生而不受管制。膀胱是貯存尿液的中空器官，主要由肌肉構成，位於下腹部，其內部接觸尿液的表面覆有一層防水的移形上皮細胞組織，最常見的膀胱癌大多源自於這一層移形性上皮細胞，稱為膀胱移行上皮細胞癌(Transitional cell carcinoma of urinary bladder)或稱為膀胱泌尿上皮細胞癌(Urothelial cell carcinoma of urinary bladder)。膀胱癌不管是發生率或死亡率都有逐年上昇之驅勢。其五年平均存活率為六成左右，因為膀胱癌常反覆地再發，所以病程延長，而且病人需經常出入醫院，因此生活品質不佳。
當第一線可治療高危險性膀胱癌的方法失敗時，其存活率是堪慮的。由於大部份的膀胱癌都屬於表淺性的膀胱癌，雖然可以反覆地施行經由尿道切除膀胱癌手術以及施行膀胱內化學藥物和免疫藥物的灌藥治療，還是會有膀胱癌的復發需要去治療，對於這些侵入性的治療，都會併發它們的副作用，影響病人的生活品質。在另一方面細胞凋亡已經被證實可以維持基本生理平衡的細胞活性，它也是一種免疫防禦的機制，藉由排除這些被誘發破壞或過多不正常的增生細胞，扮演著一種抗癌作用以及保護機轉的角色。已有證明顯示某些化學物質的抗癌作用較好的方法之一就是誘導細胞凋亡。
在人類食物的蔬菜水果中，被發現含有豐富的Chalcone可以預防癌症。在這個研究中，發現Chalcone(1,3-Diphenyl-2-propenone)在人類膀胱癌T24和HT1376兩個細胞株中有化學預防的作用，結果顯示Chalcone可以抑制T24和HT1376細胞的增生，以及引誘細胞凋亡和阻礙細胞週期的進行，使其停止在G2/M期。西方墨點分析試驗顯示Chalcone有意義的增加p21和p27蛋白質的表現，以及降低cyclin B1，cyclin A和Cdc2的表現來促成細胞週期的停止。另外Chalcone可以增加Bax和Bak的表現，及降低Bcl-2和Bcl-XL的表現，接著激發了粒線體細胞凋亡途徑(釋放cytochrome c和活化caspase-9 和caspase-3)。結果顯示Chalcone在人類膀胱癌T24和HT1376細胞中的增生抑制作用上，以及在誘發粒腺體細胞凋亡途徑和抑制NF-κB存活系統上將扮演重要的角色。

Chalcones are cancer preventive food components found in a human diet rich in fruits and vegetables. In this study, we first report the chemopreventive effect of chalcone (1,3-Diphenyl-2-propenone) in two human bladder cancer cell lines, T24 and HT-1376. The results show that chalcone inhibits the proliferation of T24 and HT-1376 cells by inducing apoptosis and blocking cell cycle progression in the G2/M phase. Western blot assay showed that chalcone significantly increases the expression of p21 and p27 proteins, and decreases the levels of cyclin B1, cyclin A and Cdc2, thereby contributing to cell cycle arrest. In addition, chalcone increased the expression of Bax and Bak, but decreased the levels of Bcl-2 and Bcl-XL and subsequently triggered mitochondrial apoptotic pathway (release of cytochrome c and activation of caspase-9 and caspase-3). Our study suggests that the induction of mitochondrial pathway and inhibition of the NF-κB survival system may play important roles in the antiproliferative activity of chalcone in T24 and HT-1376 cells.

中文摘要…………………………………………………………………………I
英文摘要…………………………………………………………………..…...III
本文目錄…………………………………………………………………...…..IV
圖表目錄………………………………………………………………...…….VII
縮寫表………………………………………………………………...….......VIII

第一章、緒論………………………………………………………………...…1
1-1 膀胱癌的流行病學研究(膀胱癌形成的原因) ……………………..1
1-2 膀胱癌的症狀………………………………………………….....….2
1-3 膀胱癌的診斷及分期……………………………………….....….....3
1-4 膀胱癌的治療…………………………………………………..........4
1-5 預後及追蹤………………………………………………….......…...5
1-6 膀胱癌的分子致癌機轉……………………………………......……6
1-7 Chalcone處理膀胱癌症的研究動機及策略…………………….….9
第二章、材料與方法……………………………………………………….....15
2-1 材料（試劑）……………………………………………...….....…15
2-2 儀器設備............................................................................................15
2-3 細胞培養(cell culture)……………………………...…………........16
2-4 細胞增生分析(Cell proliferation assay)............................................17
2-5 細胞週期分析(Cell cycle analysis)...................................................18
2-6 細胞凋亡的測量(Measurement of apoptosis)...................................18
2-7 Caspase-9和caspase-3的活性分析(Assay for caspase-9 and caspase-3 activity)…………………………………………….....…20
2-8 分離細胞中的粒線體(Mitochondria)..……..…………………..….21
2-9 分離細胞質與細胞核…………………………………………........22
2-10 NF-κB的活性(The activity of NF-κB)…………………………...…22
2-11 準備質體(NF-κB SEAP plasmid)……………………………..........23
2-12 利用SEAP(Alkaline phosphatase)測定NF-κB的活性………....…24
2-13 西方墨點法分析(Western blot assay)……………………..…...…..24
2-14 統計分析(Statistical analysis)………………………………….......25
第三章、結果……………………………………………………………..…...26
3-1 Chalcone在T24和HT-1376細胞的抗增生作用(Antiproliferative effect of chalcone in T24 and HT-1376 cells)………………...…....26
3-2 Chalcone在細胞週期分佈和細胞凋亡誘導的作用(Effects of chalcone on cell cycle distribution and apoptosis induction)….…...26
3-3 Chalcone在細胞週期相關蛋白質表現的作用(Effect of chalcone on the expression of cell cycle-related proteins)……………...….…....27
3-4 Chalcone在粒線體細胞凋亡途徑表現的作用(Effect of chalcone on the expression of mitochondrial apoptotic pathway)……………….28
3-5 Chalcone抑制NF-κB轉位和活性(Chalcone inhibited the translocation and activation of NF-κB).............................................29
第四章、討論……………………………………………………………….....30
第五章、結論……………………………………………….............................33
參考文獻……………………………………………………………………….34
圖表…………………………………………………………….........................44
附錄…………………………………………………………….........................58

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