Topoisomerase II-mediated alterations of K562 drug resistant sublines

In order to further elucidate the roles of DNA topoisomerase II (topo II) subtypes, alpha and beta, as drug targets in chemotherapy, we have determined the enzyme levels in K562 cells selected for resistance to mitoxantrone (K562/Mxn), daunorubicin (K562/Dnr) and idarubicin (K562/lda 20 and K562/Ida 60), as well as topo II-DNA complex formation, DNA damage and cytotoxicity, induced by topo II interactive agents, for example etoposide, teniposide, mitoxantrone and amsacrine. As compared to the parental cells, topo II alpha/beta protein levels in K562/Mxn, K562/Dnr, K562/Ida 20 and 60 lines, measured with Western blot, were 17/67%, 85/88, 24/31% and 10/7% respectively. DNA damage, determined by DNA unwinding technique, induced by teniposide and amsacrine correlated with both topo II alpha/beta protein levels (r(2) = 0.8/0.9, P = 0.03/0.01 and r(2) = 0.8/0.9, P = 0.04/0.01, respectively). Topo II-DNA complex formation induced by all studied drugs correlated with topo II beta protein levels (r(2)-range 0.8-0.9, P-range 0.01-0.04), while the correlation with topo II alpha was weaker. Topo II alpha/beta protein levels tended to show an inverse correlation with the cytotoxicity of etoposide (r(2) = - 0.9/ - 0.7, P = 0.01/0.06). The overall topo II-DNA complex formation correlated with drug-induced DNA damage (r(2) = 0.9, P= 0.0001), whilst not with the cytotoxicity. Our findings indicate that both topo II isozymes are the targets of the antitumor agents studied, and of potential clinical relevance for prediction of treatment efficacy. They could play a role in tailored chemotherapy.