Role of proliferation in determining sensitivity to topoisomerase II-active chemotherapy agents

Abstract

We have examined the relationship between topoisomerase II content and the DNA cleavage activity and cytotoxicity of etoposide during proliferative and quiescent culture conditions. In proliferating cultures of Chinese hamster ovary (CHO) cells, human lymphoblastic CCRF cells, and mouse leukemia L1210 cells, there was easily detectable topoisomerase II by immunoblotting. In contrast, quiescent CHO cells contained virtually no detectable topoisomerase II, while the content of L1210 cells was unchanged. Enzyme content of quiescent CCRF cells was diminished but detectable. DNA cleavage activity induced by etoposide correlated well with enzyme content in proliferating and quiescent cells. Quiescent CHO and CCRF cultures were highly resistant to the cytotoxic effects of etoposide as expected. However, despite unchanged enzyme content and DNA cleavage activity, there was also significant resistance observed in plateau L1210 cells. We have also investigated topoisomerase content and drug activity as a function of cell cycle progression. Following serum stimulation of confluent BalbC/3T3 cells, maximal etoposide-induced DNA cleavage activity is observed in G2/M and is associated with an increase in topoisomerase II content. Maximum cytotoxicity, however, occurs during mid to late S phase. Our data suggest that topoisomerase II content may be an important determinant of chemotherapeutic sensitivity during alterations in the proliferative status of the cell. However, it is clear that other factors must be involved in cell sensitivity, and elucidation of these may contribute to our understanding of the mechanism of action of these drugs.