Nakata M, Ueda K, Urata Y, Konishi E, Kishimoto M, Azumi Y, Itoi H, Yamagishi H, Oka T, Ashihara T
First Dept. of Pathology, Kyoto Prefectural University of Medicine, Japan.
Gan To Kagaku Ryoho. 1991 Mar;18(3):383-6.
In situ nick translation (ISNT) is a method to detect DNA single strand break (nick) at each cellular level. In this study, peplomycin (PEP)-induced DNA strand breaks and its repair were examined using non-radioactive ISNT method. Human fibroblasts were cultured with various concentrations of PEP for different durations, and then DNA strand breaks were evaluated by ISNT method. The signal intensity of DNA strand breaks of the PEP-treated fibroblasts was increased in a dose dependent manner. At the fixed concentration of PEP, it reached to a maximum level after 10 min. of culture and remained unchanged for at least 24 hours. In addition, DNA strand breaks induced by PEP was rapidly repaired in 10 min. after washing. Thus, the non-radioactive ISNT is thought to be a quick, sensitive and specific method not only to detect DNA strand breaks but also to observe its repair.
原位缺口平移（ISNT）是一种在细胞水平检测DNA单链断裂（缺口）的方法。在本研究中，使用非放射性ISNT方法检测了培普利霉素（PEP）诱导的DNA链断裂及其修复情况。将人成纤维细胞用不同浓度的PEP培养不同时间，然后用ISNT方法评估DNA链断裂情况。PEP处理的成纤维细胞的DNA链断裂信号强度呈剂量依赖性增加。在固定的PEP浓度下，培养10分钟后达到最高水平，并至少保持24小时不变。此外，PEP诱导的DNA链断裂在洗涤后10分钟内迅速修复。因此，非放射性ISNT被认为是一种快速、灵敏且特异的方法，不仅可用于检测DNA链断裂，还可用于观察其修复情况。
