The DNA demethylating 5-azaC induces endoreduplication in cultured Chinese hamster cells

We have investigated the possible influence of 5-azacytidine (5-azaC) substitution for cytidine into DNA on topoisomerase II (topo II) function in chromosome segregation. The endpoint chosen has been the induction of endoreduplicated cells at mitosis showing diplochromosomes. Experiments were performed in the presence and absence of the cytidine analogue to assess the degree of 5-azaC-induced DNA hypomethylation, using differential cutting by restriction endonucleases Hpa II and Msp I. Using the pulsed-field gel electrophoresis (PFGE) technique, we have also observed a protective effect provided by 5-azaC treatment against DNA breakage induced by the topo II poison m-AMSA. Concentrations of 5-azaC shown as able to induce extensive DNA hypomethylation and capable to protect DNA from double-strand breaks induced by m-AMSA were used for our cytogenetic experiments to analyze chromosome segregation. Our results seem to indicate that the presence of 5-azaC in DNA induces a dose-dependent increase in the yield of endoreduplicated cells that parallels the levels of hypomethylation observed.