Cytogenetic assay in apoptosis investigations

The mitotic figure, named premature anaphase (PA) or C-anaphase, could be considered as a cytogenetic forerunner of following cell apoptosis in G1 phase. To confirm this hypothesis the comparative analysis was performed using cytogenetic, cytologic, flow cytometric and DNA fragmentation methods upon the cells with different proliferative ability and degree of differentiation. PA level was significantly increased in bone marrow and blood cells in vitro in cases of acute lymphoblastic leukemia and non-Hodgkin lymphoma, decreased until total disappearance in remission and not revealed in control. Particularly high PA level was registered in embryonal liver's haemopoetic stem cells ex vivo. Flow cytometry measurements showed appearance of additional sub-G1 peak of apoptotic DNA loss both in leukaemic and embryonal cells, whereas DNA-ladder phenomenon was revealed just only in embryonal samples in vivo. Significant positive correlation between the frequency of cells with apoptotic DNA loss and PA level on the chromosomal preparations was registered. Thus, premature anaphase phenomenon is considered as non-random event, associated with high risk of following cell death.