- 學位論文
使用彗星修復能力試驗評估個體間DNA修復能力的差異
Inter-individual difference in DNA repair capacity by Comet DNA repair kinetic assay
摘要
因為DNA修復具有多種路徑且彼此具有相互支援的功能，而其中有多種基因與DNA修復路徑相關。且DNA修復能力與修復基因之基因多型性缺少功能性相關，故發展功能性試驗去評估影響DNA修復能力的相關因子變得越來越重要。我們以44個學生為研究對象，採集新鮮的血液樣本分離出淋巴細胞，藉由紫外光（254nm）照射使之產生DNA損傷後，在0、10、30、60分鐘的修復後，以慧星修復能力試驗測量淋巴細胞的DNA修復能力。首先以彗星修復能力試驗測試同一受試者在不同時間施測對修復能力的影響，發現DNA修復能力會因為收集時間的不同而有不同的結果。比起女性，男性具較高的基礎DNA損傷程度。以殘餘損傷百分比（%RD）與修復百分比（%RC）為修復指標，男性相對於女性同樣的也顯示出較少的殘餘損傷百分比，且男性在60分鐘內其單位時間殘餘損傷遞減率與單位時間修復率較快，此結果顯示男性修復較女性為快。這些結果指出在評估個體對於環境暴露的易感受性的研究中，測量DNA在細胞照射紫外光後的損傷修復能力是有用的工具。
並列摘要
There are multiple genes involved in DNA repairs pathways. In light of the presence of multiple DNA repair enzymes with back-up functions and the lack of functional relevance to the genetic polymorphisms of DNA repair genes, it has become more and more important to apply functional assay to address environmental and occupational health concerns. A modified challenge mutagen sensitivity assay, Comet DNA repair kinetic assay, was applied to characterize the DNA repair capacity and to investigate the factors that modify individual DNA repair capacity. The study population consisted of 44 students, aged from 20 to 29 years. Fresh blood samples were irradiated with UV (254 nm) and the kinetics of DNA repair in lymphocytes assessed by comet assay 0, 10, 30 and 60 min after irradiation. To assess the intraindividual variation in DNA capacity, the DNA repair capacities were found not to track from time to time (intraclass correlation coefficient). A higher basal damage was found in male students. By measuring the percentage of residual DNA damage (%RD) and percentage of repair capacity (%RC) , male also show less amount of residual damage (%RD) shortly after irradiation, compared with female. By measuring residual DNA damage (%RD) per unit time and repair capacity (%RC) per unit time, the results indicate that male shorts are cyrible of repairing damage DNA faster then female student. These results indicate that the repair capacity and the DNA damage after in vitro irradiation can be a useful tool in the study of host susceptibility to environmental exposure.