Determination of DNA topoisomerase II activity from L1210 cells--a target for screening antitumor agents.
DNA topoisomerase II was isolated from mouse leukemia L1210 cells and the activity was determined by using P4 phage knotted DNA and pBR 322 DNA as the substrates. Based on these results, a method for screening antitumor agents by using DNA topoisomerase II as a target was established. The experiments showed that DNA topoisomerase II catalyzed pBR 322 DNA breaking and relaxing which were reversible and dependent on ATP. The activity was increased 2-4 times in the presence of ATP 1 mmol.L-1. In contrast with type II enzyme, the activity of DNA topoisomerase I was completely inhibited in the presence of ATP 1 mmol.L-1 and had full activity in the absence of ATP. Type II enzyme also showed the unknotting activity by using p4 phage knotted DNA as a substrate. DNA cleavage and relaxing reaction induced by type II enzyme increased 5-fold in the presence of Doxorubicin (Dox) 1 microgram.ml-1 or daunorubicin (Dau). Etoposide (Eto) and aclarubicin B (Acl B) also stimulated the reaction at 100 micrograms.ml-1. The cleavage reaction resulted from topoisomerase II was inhibited by other agents, such as frankincense extracts, terpenic compounds (BC series).
Wang
L G
LG
Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing.
Liu
X M
XM
Ji
X J
XJ
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D004249:DNA Damage; D004250:DNA Topoisomerases, Type II / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D004357:Drug Synergism; D007939:Leukemia L1210 / Q000201:enzymology* / Q000473:pathology; D051379:Mice; D008811:Mice, Inbred DBA; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000970:Antineoplastic Agents; D004250:DNA Topoisomerases, Type II