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研究生: 林明興
Ming-Hsing Lin
論文名稱: 不同型式鍺化物對細胞生長與輻射敏感性的效應之比較性研究
Effect of different forms of germanium compounds on cell growth and radiosensitivity
指導教授: 林立元
Lih-Yuan Lin
口試委員:
學位類別: 碩士
Master
系所名稱: 生命科學暨醫學院 - 分子與細胞生物研究所
Institute of Molecular and Cellular Biology
論文出版年: 2006
畢業學年度: 94
語文別: 中文
論文頁數: 58
中文關鍵詞: 細胞週期輻射敏感性
外文關鍵詞: germanium, cell cycle, radiosensitivity
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  • 本篇論文主旨在於比較三種不同型式的鍺化物—氧化鍺(GeO2)、有機鍺(Ge-132)以及奈米鍺顆粒—對細胞生長與輻射敏感性等效應的差異。將中國倉鼠卵巢細胞(CHO K1)以5 mM Ge-132持續處理48小時,並不會影響其生長速率;相反的,處理5 mM GeO2的細胞則明顯可見生長速率遲緩。造成生長遲緩的原因主要是由於細胞週期的停滯。我們發現經過氧化鍺及奈米鍺顆粒處理過的細胞,其細胞週期分佈會停留在G2/M期,而有機鍺並不會造成這樣的情況。此外,氧化鍺及奈米鍺顆粒亦可增加細胞的輻射敏感性,有機鍺仍然無此現象。經過分析,細胞對有機鍺的攝取速率低於氧化鍺,在同樣的處理時間內,細胞內累積的有機鍺比氧化鍺來得少。但即使讓細胞攝取了相同量的鍺,有機鍺和氧化鍺的效應仍舊不同。氧化鍺及奈米鍺是透過增加輻射線對DNA的傷害並延緩DNA傷害的修補以提升細胞的輻射敏感性。我們發現氧化鍺和奈米鍺處理過的細胞,在經過X射線照射之後,會延長細胞週期停留在G2/M期的時間。而以鹼性彗星檢測及西方墨點法偵測□-H2AX的表現量,也可觀察到處理氧化鍺和奈米鍺的細胞,在經過X射線照射後,其DNA斷裂的情況加劇;若使細胞繼續培養4小時,DNA傷害的情況也尚未回復,也因此導致細胞的輻射敏感性增加。


    We compared the cellular responses to three different forms of germanium compounds, i.e. inorganic-Ge (germanium oxide, GeO2), organic-Ge (Ge-132), and nano-Ge particle. Treating Chinese hamster ovary K1 cells with 5 mM Ge-132 did not alter the growth rate as compared to those of untreated cells within 48 hours. On the contrary, 5 mM GeO2 treatment delayed the cell growth through arresting the cells at G2/M phase. The same effect was observed in cells treated with nano-Ge. However, the population of cells in G2/M phase remained unchanged when receiving Ge-132. Moreover, treating cells with GeO2 and nano-Ge enhanced radiosensitivity, whereas the effect was not observed in Ge-132 treated cells. The cellular uptake rate of Ge-132 was slower than that of GeO2, which resulted in a less accumulation of Ge content in Ge-132 treated cells. However, the Ge content was similar when cell received 5 mM Ge-132 or 1 mM GeO2 for 12 hours. In this condition, a different radiosensitizing effect was still observed. GeO2 and nano-Ge enhanced radiosensitivity through elevating DNA damage as analyzed by alkaline comet assay and the quantity of □-H2AX. After exposing to the X-ray, a prolonged G2/M arrest was found in cells treated with GeO2 and nano-Ge. The presence of inorganic Ge compounds also affects the efficiency of DNA repair after X-irradiation. Enhancement of DNA break and reduction of DNA repair efficiency concurrently cause the increase of radiosensitivity.

    目錄 謝誌 i 中文摘要 ii 英文摘要 iii 目錄 iv 圖目錄 v 第一章 緒論 1 一、鍺化物的簡介 1 二、細胞週期調控 5 三、DNA雙股斷裂之傷害與修補 7 四、研究目的 9 第二章 材料與方法 11 一、 化學藥品 11 二、 細胞株、細胞培養 11 三、 細胞生長速率的測定 11 四、 細胞週期分佈的測定 12 五、 X光照射(X-irradiation) 12 六、 細胞群落形成分析(Clonogenic survival assay) 13 七、 細胞內鍺含量的測定 13 八、 奈米鍺顆粒溶解度測試 14 九、 鹼性彗星檢測(Alkaline comet assay) 14 十、 細胞全蛋白質的萃取 16 十一、 蛋白質濃度的測定 16 十二、 西方墨點法(Western blotting) 17 十三、 數據分析 17 第三章 結果 18 一、不同型式鍺化物對細胞生理的影響 18 二、提升輻射敏感性的可能機制探討與比較 23 三、結論 27 第四章 討論 28 參考文獻 34 附圖 41 附錄 58 圖目錄 圖一、有機與無機鍺化合物對細胞生長的影響 41 圖二、有機與無機鍺化合物對細胞週期運行的影響 43 圖三、有機或無機鍺化合物對細胞輻射敏感性的效應 44 圖四、細胞對有機和無機鍺化合物攝取量的檢測 45 圖五、細胞對不同濃度有機和無機鍺化合物的攝取量差異 46 圖六、細胞含有相似含量的有機和無機鍺化合物時所表現出的輻射敏感性 47 圖七、不同粒徑奈米鍺顆粒的溶解度測試 48 圖八、奈米鍺顆粒對細胞週期運行的影響 49 圖九、奈米鍺顆粒對細胞輻射敏感性的效應 50 圖十、以不同型式鍺化物處理細胞並照射輻射線後,其細胞週期分佈的改變 51 圖十一、不同型式鍺化物在增進輻射線對DNA傷害的差異 53 圖十二、不同型式鍺化物對延緩DNA傷害修補的差異 55 圖十三、奈米鍺顆粒增進細胞照射輻射線後DNA雙股斷裂的產生 57

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