title: Use of DBD-FISH for the study of cervical cancer progression.
abstract: DNA breakage detection-fluorescence in situ hybridization (DBD-FISH) is a procedure to detect and quantify DNA breaks in single cells, either in the whole genome or within specific DNA sequences. This methodology combines microgel embedding of cells and DNA unwinding procedures with the power of FISH coupled to digital image analysis. Cells trapped within an agarose matrix are lysed and immersed in an alkaline unwinding solution that produces single-stranded DNA motifs beginning at the ends of internal DNA strand breaks. After neutralization, the microgel is dehydrated and the cells are incubated with fluorescently labeled DNA probes. The amount of hybridized probe at a target sequence correlates with the amount of single-stranded DNA generated during the unwinding step, which is in turn proportional to the degree of local DNA breakage. A general view of the technique is provided, emphasizing its versatility for evaluating the association between DNA damage and progressive stages of cervical neoplasia.
authors.affiliation: Department of Genetics, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social (IMSS), C. P. 64720, Monterrey, Nuevo León, Mexico, elvacortes@cibinmty.net.
authors.forename: Elva I
authors.initials: EI
authors.lastname: Cortés-Gutiérrez
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Fernández
authors.forename: Martha I
authors.initials: MI
authors.lastname: Dávila-Rodríguez
authors.forename: Carmen
authors.initials: C
authors.lastname: López-Fernández
authors.forename: Jaime
authors.initials: J
authors.lastname: Gosálvez
mesh_terms: D053960:DNA Breaks; D004273:DNA, Neoplasm / Q000378:metabolism; D018450:Disease Progression; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D005453:Fluorescence; D006801:Humans; D007091:Image Processing, Computer-Assisted; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D012372:ROC Curve; D013048:Specimen Handling; D013535:Suspensions; D002583:Uterine Cervical Neoplasms / Q000473:pathology*
chemical_list: D004273:DNA, Neoplasm; D013535:Suspensions