臺灣博碩士論文加值系統

　　膀胱癌，為常見泌尿系統癌症的一種，尤為生殖泌尿腫瘤中次常見的死因。而台灣南部的烏腳病流行區則有特別高的膀胱癌發生率。傳統上用以診斷膀胱癌的方法具侵入性且花費昂貴，因此，找出診斷膀胱癌的新方法是迫切需要的。本研究的目的是利用蛋白質體學分析方法建立膀胱癌病人尿液中的蛋白質分布圖譜。選取來自台灣西南地區的膀胱癌病人、烏腳病流行區的膀胱癌病人以及對照組共三組配對成實驗組進行尿液中蛋白質體內容的分析。人體的體液如尿液，具有複雜的基質；其中可能包含各種無機鹽類、緩衝液及蛋白質，而這些成分可能對樣本在質譜的分析上造成干擾甚或失敗。雖然蛋白質體二維膠電泳技術已發展多年，但於研究過程中發現，其對於尿液蛋白質體的鑑定工作上仍有限制。因此，本研究所使用的方法是將尿液淨化後以奈米高效能液相層析電噴灑串聯式質譜儀來分析尿液中蛋白質體。此分析方法的建立工作將有利於往後以蛋白質體資料庫鑑定膀胱癌病人蛋白質體之不同表現，進一步甚可尋找膀胱癌的蛋白質生物指標。

　Bladder cancer, one of the common urologic cancers, represents the second most common cause of death among genitourinary tumors. The higher bladder cancer incidence rates have been observed in the endemic area of black foot disease (BFD) in southern Taiwan. The traditional bladder cancer diagnose methods are both invasive and expensive, thus developing new approaches for the diagnosis of bladder cancer are in urgent need. The aim of this study is utilized proteomic approach to establish urinary protein patterns. In this study, the result details protein profiles in urine from control, BFD bladder cancer, and non-BFD bladder cancer patients. Biofluids, such as urine, from very complex matrixes often contain large kinds of inorganic salt, buffers, and proteins. However, the presence of contaminants virtually ensures the failure of subsequent analysis of the sample by mass spectrometry. Although impressive improvements in 2DE technologies have occurred in recent years, identification of urine proteins is still challenging. In this experiment, it tends to complete a study for a urine sample cleanup strategy and identification by using nano-HPLC-ESI-MS/MS spectrometry. These proteins are valuable for mass identification of differentially expressed proteins involved in proteomics database and screening biomarker to further study in human bladder cancer. The significance for creating a strategy of urinary proteome analyze for new bladder cancer marker proteins discover, and for their simultaneous display and analysis in patients suffering from bladder disorders has been examined.

中文摘要…………………………………………………………………………I
英文摘要…………………………………………………………………………II
謝誌………………………………………………………………………………III
主目錄……………………………………………………………………………V
表目錄……………………………………………………………………………VII
圖目錄……………………………………………………………………………VIII
中英文對照表……………………………………………………………………IX
英文縮寫對照表…………………………………………………………………X

第一章 緒論
第一節 研究動機…………………………………………………………1
第二節 研究目的…………………………………………………………1
第二章 文獻探討
第一節 膀胱癌……………………………………………………………2
第二節 蛋白質體學………………………………………………………10
第三節 尿液蛋白質體分析………………………………………………15
第三章 研究材料與方法
第一節 研究架構…………………………………………………………22
第二節 尿液樣品採取……………………………………………………23
第三節 樣品前處理………………………………………………………24
第四節 蛋白質二維膠體電泳分析………………………………………27
第五節 高效能液相層析質譜儀分析……………………………………35

第四章 結果與討論
第一節 蛋白質二維膠體電泳分析結果…………………………………42
第二節 高效能液相層析串聯質譜儀分析結果…………………………53
第五章 結論
第一節 結論………………………………………………………………64

參考文獻…………………………………………………………………………66
附錄一 重複分析樣品的三次分析base peak層析圖………………………74
附錄二 解凍/冷凍樣品的三次分析base peak層析圖…………………….75
附錄三 樣品的三次分析base peak層析圖…………………………………76
附錄四 健康尿液中蛋白質體……………………………………………….78
附錄五 烏腳病流行區膀胱癌特異蛋白胺基酸序列……………………….82

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