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《Chinese Journal of Cancer》 1991-03
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DETERMINATION OF DNA TOPOISOM ERASE II FROM MOUSE LEUKEMIA L1210 CELLS A TARGET FOR SCREENING ANTITUMOR AGENTS

LUI Xiao-mei, WANG Long-gui and JI Xiu-juan (Institute of Materia Medica, Chinese Academy of Medical Scienes, Beijing 1000650)  
DNA topoisomerases play very important roles in DNA breakage, reunion replication, and other genetic processes. Recently it has been found that DNA topoi-somerases I and II are fruitful targets for both antimicrobial and oncologic drug development. In the present work a new method for screening antitumor agents by using DNA top-oisomerase II as a target has been established according to the phenomena that the enzyme activity can be affectd by some types of antitumor agent.Based on the method reported by Sullivan et al. with some modification, the DNA top-oisomerase II was extracted from mouse leukemia L1210 cells. The experiments have shown that this enzyme can catalyze pBR 322 DNA breaking and relaxing which are revesible and can be inhibited by high concentration of sodium chloride. Experiments also showed that p-BR DNA cleavage induced by the enzyme can be stimulated by some antitumor drugs. The cleavage reaction activity increased fivefold in the presence of 1 ug/ml of ADM or DNR. Vp-16 and ACM-B can also stimulated the reaction in the higher concentration. The results, however, also indicated that the cleavage reaction resulted from topoisomerase II can be inhibited by other antitumor agents, such as herb ectracts, BC series.
【Key Words】： DNA topoisomerase Ⅱ Screening method Electrophoresis p BR 322 DNA L1210 cells.
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