摘要

在許多研究中，重荷電粒子已被證明會造成較複雜的 DNA 傷害
(complex DNA damage)，而且此種傷害將會造成較顯著的生物效應。
然而，關於重荷電粒子引起 DNA 傷害的修復機制仍有待探討。因此，
本實驗利用國立清華大學加速器實驗室的重荷電粒子生物照射系統進
行細胞照射，並分析 γ-h2ax 蛋白的表現量，藉以探討重荷電粒子引起
DNA 傷害的修復動態。
本實驗使用 2 MeV α 粒子對 HeLa 細胞進行照射，照射劑量分別為
10、20、50、100 α粒子/細胞。照射完畢後，於不同修復時間點對細胞
樣品進行固定 (fixed) 以及 γ-h2ax 螢光免疫染色，隨後使用反向式螢光
顯微鏡 (TE2000, Nikon) 擷取 γ-h2ax 螢光影像。最後，將取得影像載入
image-j 軟體並分析不同修復時間 γ-h2ax 蛋白表現量，以獲得 DNA 修復
反應動態。
結果顯示，細胞於照射後 5 分鐘，已可觀察到 γ-h2ax 聚集，且 γ-h2ax
蛋白表現量隨著時間增加而逐漸上升，並於 20 ~ 30 分鐘左右達最大值。
而γ-h2ax 累積速率，隨著劑量增加而上升。在達到最大值後，γ-h2ax 的
表現量隨著時間增加而緩慢減少，並且於 48 小時後仍殘留相當高的比
率 ( > 25 %)，其殘留比率亦隨著劑量增加而上升。雙指數衰減擬合結果
顯示，代表單一 DNA 傷害 (simple DNA damage) 修復的成分隨著劑量
上升而減少，而代表複雜 DNA 傷害 (complex DNA damages) 修復的成
分則隨劑量上升而增加。此一部分結果顯示，隨著重荷電粒子劑量增加，
將造成複雜 DNA 傷害的比率增加。
於本論文中，我們發現細胞對於 DNA 傷害的修復能力將會依照重
荷電粒子劑量增加而改變。隨著劑量上升，使 DNA 傷害的複雜程度增
加，造成細胞對於 DNA 傷害的修復能力下降，進而產生更顯著之生物
效應。

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