臺灣博碩士論文加值系統

根據衛生署的台灣地區癌症登記調查報告，膀胱癌佔了泌尿系統癌症之大部份，而且有逐年增加的趨勢。根據民國九十四年統計，台灣九十四年度男性因膀胱癌死亡人數為507人，女性為240人，死亡率在各部位癌症中排行第十五，平均每十萬人有三人死於膀胱癌。目前在膀胱癌的診斷及治療上，皆缺乏有效準確的生物指標得以協助治療策略的制定。膀胱癌發展的分子機轉，目前所知仍相當有限。因此之故，在基礎研究上進一步研發有效而方便的診斷預後指標是各方研究者共同的理想。隨著晶片技術的發展，利用晶片平台對許多不同的檢體或實驗材料進行測試比對，所得到大量的資料以作為研究方向的參考或分析趨勢，已蔚為風潮，並成為非常寶貴的研究依據。我們利用cDNA晶片的篩選技術，比較正常的膀胱上皮細胞與早期膀胱癌細胞株RT4的基因表現。經過資料整理及文獻蒐集後，從中挑選出25個基因做進一步的研究。我們利用反轉錄聚合酶鏈式反應(RT-PCR)的方式，衡量各基因在數種膀胱癌細胞株的基因表現情形，再從中進一步挑選基因做更進一步的研究。其中內皮素Ⅰ (Endothelin 1)便是其中之一的致癌潛力基因。內皮素Ⅰ過去曾經被報導與數種癌症進程相關，但與膀胱癌相關的研究資料相當缺乏。在我們的實驗中，雖然外加內皮素Ⅰ對膀胱癌細胞株的生長情形沒有明顯促進的效果;但是加入內皮素Ⅰ的拮抗劑BQ123 (cyclo(-D-Trp-D-Asp2- Pro-D-Val-Leu-)•Na+)，對於膀胱癌細胞的生長卻有抑制的情形出現。另外，加入內皮素Ⅰ可以促進表皮細胞生長因子接受體(EGFR)的磷酸化作用，暗示著內皮素Ⅰ可能透過活化細胞中訊息傳遞以調節細胞的功能。然而加入內皮素Ⅰ拮抗劑後，細胞型態有傾向細胞凋亡的情形。根據我們的研究結果顯示，內皮素Ⅰ有利於細胞的生存，並且極可能可以影響膀胱癌發展過程。因此，內皮素Ⅰ拮抗劑值得深入研究做為治療膀胱癌的藥物選擇之一。

Bladder cancer is one of the most common urinary tract malignancies in Taiwan. Bladder tumors are predominantly transitional cell carcinoma (TCC), but display significant variation in clinical behavior, propensity to recur, progression, and prognosis, which likely reflect genetic heterogeneity. Thus, an improvement of tumor diagnosis or prognostic prediction is mandatory in the determination of appropriate treatment planning for cancer patients. Thus, identification of genes associated with pathogenesis of bladder cancer may be of great help in cancer therapy and prognostic prediction. In this pilot study, cDNA microarray screening was first utilized to search for differential expression gene lists between normal bladder epithelium and RT4 cancer cell line. Then, gene ontology was analyzed by NCBI database mining. Those genes thought to have potential relevance were screened by RT-PCR (n = 25), and then confirmed by real-time PCR. By using this strategy, we demonstrated that endothelin 1 (ET-1) is one of the candidate genes involved in the carcinogenesis of human bladder. ET-1 was universally expressed in all uroepithelial cell lines tested. The cognate receptor, ETAR (endothelin receptor A) was also expressed in all cancer cell lines, while only J82 cells expressed the endothelin receptor B. The results suggest that ET axis may play a role in the pathogenesis of bladder cancer. To verify our hypothesis, The ETAR antagonist (BQ123) was treated on several bladder cancer cell lines. We found that treatment with BQ123 successfully suppressed the proliferation of tumor cells. However, treatment with ET-1 produced slight growth stimulatory effect on bladder cancer cells. A similar result was also observed in ET-2 or ET-3. Interestingly, ET-1 treatment could activate phosphorylation of EGFR in TSGH8301 cancer cells, implying a cross-talk between ET axis and EGFR signaling pathway. When cancer cells were treated with BQ123, tumor cells tended to have larger nuclei and increased apoptotic bodies. Our data suggest that ET axis may play a role in bladder carcinogenesis, and activation of its signaling pathway might provide growth advantage for human bladder cancer cells. The clinical significance of our observation deserves further investigation.

1. Abstract in English -3-
2. Abstract in Chinese -5-
3. Acknowledgements -7-
4. Contents -8-
5. Index of Tables -9-
6. Index of Figures -10-
7. Abbreviations -11-
8. Introduction 1
9. Materials and Methods 4
10. Results 9
11. Discussion 14
12. References 17
13. Appendix of Tables 20
14. Appendix of Figures 27

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