Analyzing Topoisomerase II Cleavage Complexes Using Flow Cytometry

Flow cytometry is a powerful technique used in biology and medicine to analyze and quantify characteristics of individual cells in a heterogeneous population. It provides information about various cellular properties, such as size, complexity, surface markers, and internal components. We developed a flow cytometry-based method for the analysis of drug-induced topoisomerase II cleavage complex (Top2cc). This method allows a rapid analysis of a high number of cells in their cell cycle phase context. Moreover, it can be applied to almost any human cell type, including clinical samples. The methodology is valuable for conducting high-throughput analyses of drugs that target Top2, enabling the discrimination of the specific isoform affected, and tracking its removal. Since drug-stabilized Top2cc may interfere with several nuclear processes of DNA, this method allows the study of different pathways that may be hampered, evaluating their impact under different cellular conditions.