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研究生:王裕明
研究生(外文):Yu-Ming Wang
論文名稱:攝護腺癌化時之蛋白質變化
論文名稱(外文):Protein expression in prostate cancer progress
指導教授:黃弘文黃弘文引用關係
學位類別:碩士
校院名稱:國立中山大學
系所名稱:生物醫學科學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:75
中文關鍵詞:攝護腺二維電泳
外文關鍵詞:prostatetwo-dimensional electrophoresis
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攝護腺癌在年老時常見的惡性腫瘤之一,攝護腺特異性抗原(prostate specific antigen, PSA)是目前最重要用來早期檢測攝護腺癌的血清腫瘤標誌,但PSA本身無法偵測辨別攝護腺癌之各種型態,故發展各種攝護腺癌之分子依然為科學界所重視,本實驗利用二維電泳 (two-dimensional electrophoresis)與基質輔助雷射脫附游離飛行時間(matrix-assisted laser desorption ionization-time of flight, MALDI-TOF)質譜法來探討正常人和攝護腺癌病人血液蛋白質的差異,與比較攝護腺癌在治療過程中血液蛋白質的變化,我們發現在癌化過程和治療後分別共有31和17個蛋白質表現量改變,目前我們經由SWISS-PROT資料庫比對和基質輔助雷射脫附游離飛行時間方法,已確認了Fibrinogen gamma chain, Fibrinogen alpha/alpha-E chain, major histocompatibility complex, class I, C和Mayven四個蛋白質在癌化時表現量提高。Fibrinogen, MHC在癌化過程中病人血液變化的結果與最近的報告類似, 而Mayven的表現變異則僅在本研究中發現。


Prostate cancer is one of the most common malignant tumors in solid organs of old men. Prostate-specific antigen (PSA) is a valuable prostate cancer biomarker that is now wildly used for population screening, diagnosis, and monitoring of patients with prostate cancer. Howere it is reported that PSA is not feasible to discriminate the progress of prostate cancer, so many investigators still works on developing new biomarker of prostate carcinoma. Here, we propose the study of differentially expressed prostate proteins in blood of patients.
With the aid of two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption-induced time of flight (MALDI-TOF) mass spectrometry, comparison of normal men and prostate cancer patients serum proteins and analysis protein variation of different stages of prostate cancer serum. We find 31 and 17 protein spots overexpression in cancer development and after treatment, respectively.
At present, with the aid of SWISS-PORT database and MALDI-TOF mass spectrometry, we had identified fibrinogen gamma chain, fibrinogen alpha/alpha-E chain, major histocompatibility complex (MHC), class I, C and Mayven were overexpressed in prostate cancer development, where Mayven is fist reported by us.


中文摘要 I
英文摘要........................... II
縮寫.............................. III
一、攝護腺癌的介紹..................... 1
二、攝護腺癌的分期..................... 1
三、病原學............................. 2
四、已知癌症發生的可能致癌機轉......... 3
五、致癌基因和原致癌基因............... 4
六、腫瘤抑制基因....................... 4
七、男性賀爾蒙接受體................... 5
八、臨床診斷........................... 5
九、攝護腺專一抗原..................... 6
十、攝護腺專一抗原和kallikrein家族..... 7
十一、攝護腺專一抗原的分子形式......... 8
材料................................... 9
方法
實驗儀器............................... 12
實驗商業套組........................... 12
實驗方法
一、檢體來源與製備................... 12
二、蛋白質二維電泳................... 14
三、基質輔助雷射脫附游離/飛行時間質儀器. 19
四、蛋白質一維電泳................... 20
五、西方點墨法....................... 21
結果
一、血清二維電泳以Coomassie blue R350染色. 23
二、以Cibacron Blue 3GA去除血清中白蛋白.. 23
三、血清二維電泳銀染呈色和Swiss Prot 生物資訊中心血漿二維電泳銀圖........................ 23
四、正常人和攝護腺癌病人血清的二維電泳圖比較 24
五、血清二維電泳蛋白質確認................. 25
六、血清和精液PSA的測定.................... 26
七、常人和攝護腺癌病人精液以二維電泳分析比較.26
八、二維電泳分析攝護腺腫大和攝護腺癌病人之攝護腺組織蛋白質差異............................... 26
結果與討論
一、血清樣本蛋白質之二維電泳分析............ 27
二、以SWISS-PROT生物資訊中心的血漿二維電泳圖之蛋白質比較定位.................................. 27
三、攝護腺特異性抗原在SDS-PAGE上的位置...... 28
四、銀染對MALDI-TOF實驗的影響............... 29
五、病人的差異點探討........................ 30
六、差異點在組織和精液中的表現............. 31
結論
參考文獻................................... 33
圖......................................... 40
表......................................... 65


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