臺灣博碩士論文加值系統

本論文在建立DNA疫苗治療老鼠攝護腺癌的動物模型，以模擬利用DNA疫苗治療人類攝護腺癌的情形。在人類攝護腺治療中，通常以攝護腺特殊抗原(prostate-specific antigen, PSA)為基因療法之標的物，於是本論文在動物模型的抗原的選擇上，以正常老鼠攝護腺體表現的probasin蛋白質作為DNA疫苗基因療法的目標物。本實驗主題在利用基因重組技術，由老鼠攝護腺體取得probasin基因，與質體接合後，構築出可製造probasin的質體DNA (pIRESpuro-probasin)。再以此質體DNA做為DNA疫苗，用肌肉注射方式(i.m.)打入不同品系正常老鼠體內，觀察不同宿主引發的免疫反應，並且以組織切片觀察是否因為打入能產生大量自身抗原的質體，而使攝護腺組織產生發炎現象。對於未來動物模型的建構，利用轉染法(transfection) 將建構好的pIRESpuro-probasin質體植入失去表現probasin能力的老鼠攝護腺癌細胞株(TrampC-1)中，以模擬人類的攝護腺癌細胞分泌PSA的情形。
本論文成功建構出pIRESpuro-probasin質體DNA作為DNA疫苗，證明施打於動物體內並無副作用發生，並且構築了能分泌probasin的老鼠攝護腺癌細胞株(TrampC-1- pIRESp-probasin)，模擬人類攝護腺腫瘤，亦初步證實其腫瘤生成能力與原細胞株(TrampC-1)相同。未來實驗方向應朝結合probasin與細胞激素的DNA疫苗，輔助以建立anti-probasin monoclonal antibody，觀察細胞內或體內probasin蛋白質表現，以期能夠有效的建立以DNA疫苗治療老鼠攝護腺癌的模型。

目錄
1. 以probasin作為老鼠攝護腺癌DNA疫苗的效果評估
英文摘要………………………………………………………………. I
中文摘要……………………………………………………………….III
誌謝……………………………………………………………………..V
目錄…………………………………………………………………… VI
第一章 緒論
第一節 傳統癌症治療與攝護腺癌基因療法…………………….....1
第二節 免疫基因療法(Immuno-gene therapy)與DNA疫苗(DNA vaccine)對攝護腺癌治療之應用……………………….....5
第三節 小鼠攝護腺抗原probsain介紹………………………….....9
第四節 研究目的與內容……………………………………………11
第二章 材料與方法
第一節 構築pIRESpuro-probasin質體DNA………………………13
第二節 製作pIRESpuro-probasin質體DNA為DNA疫苗(vaccine)
……………………………………………………………..19
第三節 將pIRESpuro-probasin質體DNA送入細胞中……………23
第四節 以靜脈注入細胞至老鼠體內，觀察肺部轉移腫瘤細胞數..26
第三章 實驗結果
第一節 構築pIRESpuro-probasin質體DNA……………………...28
第二節 以pIRESpuro - probasin質體DNA為DNA疫苗(vaccine)，在C3H/HeN及C57BL/6J品系老鼠中的免疫反應………30
第三節 將pIRESpuro - probasin質體DNA送入細胞中………….32
第四節 比較TrampC-1-pIRESp-probasin及TrampC-1細胞以靜脈注射方式打入C57BL/6J老鼠後，肺部腫瘤細胞轉移之情形
…………………………………………..…………………..35
第四章 討論……………………………………………………………36
圖表……………………………………………………………………..43
附錄……………………………………………………………………..56

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