Abstract：
Genomic instability is an important biomarker in the progression of cervical carcinoma. DBD-FISH (DNA breakage detection-fluorescence in situ hybridization) is a sensitive method that detects strand breaks, alkali-labile sites, and incomplete DNA excision repair in cells of the cervical epithelium. This technique integrates the microgel immersion of cells from a vaginal lesion scraping and the DNA unwinding treatment with the capacity of FISH integrated into digital image analysis. Cells captured within an agarose matrix are lysed and submerged in an alkaline unwinding solution that generates single-stranded DNA motifs at the ends of internal DNA strand breaks. After neutralization, the microgel is dehydrated and the cells are incubated with DNA-labeled probes. The quantity of a hybridized probe at a target sequence corresponds to the measure of the single-stranded DNA produced during the unwinding step, which is equivalent to the degree of local DNA breakage. DNA damage does not show uniformly throughout the entire DNA of a cell; rather, it is confined to specific chromosomal sites. In this chapter, an overview of the technique is supplied, focusing on its ability for assessing the association between DNA damage in specific sequences and in the progressive stages of cervical carcinoma.
摘要：
基因组不稳定性是宫颈癌进展的重要生物标志物。DBD-FISH（DNA断裂检测-荧光原位杂交）是一种检测链断裂的灵敏方法，碱不稳定位点，宫颈上皮细胞的不完全DNA切除修复。该技术将来自阴道病变刮擦的细胞的微凝胶浸没和DNA展开处理与FISH的能力整合到数字图像分析中。将捕获在琼脂糖基质内的细胞裂解并浸没在碱性解链溶液中,所述碱性解链溶液在内部DNA链断裂的末端产生单链DNA基序。中和后,将微凝胶脱水并将细胞与DNA标记的探针一起孵育。靶序列处的杂交探针的量对应于在解链步骤期间产生的单链DNA的测量值。相当于局部DNA断裂的程度。DNA损伤不会在整个细胞的整个DNA中均匀显示;相反，它局限于特定的染色体位点。在这一章中,提供了该技术的概述，专注于评估特定序列中DNA损伤与宫颈癌进展期之间的关联的能力。
