Constantinou A, Kiguchi K, Huberman E
Biological and Medical Research Division, Argonne National Laboratory, IL 60439.
Cancer Res. 1990 May 1;50(9):2618-24.
Genistein, an in vitro inhibitor of topoisomerase II and tyrosine kinases, suppressed growth and induced differentiation in HL-205 cells, a clonal population of the human promyelocytic HL-60 leukemia cells, and in K-562-J cells, a clonal population of the human erythroid K-562 leukemia cells. Maturing HL-205 cells acquired either granulocytic or monocytic markers, namely, reactivity with the murine OKM1 monoclonal antibody, expression of nitroblue tetrazolium dye reduction, and staining for nonspecific esterase. The maturing K-562-J cells stained with benzidine, which indicates the presence of hemoglobin, an erythroid maturation marker. Although the acquisition of the maturation markers in both HL-205 and K-562-J cells was time dependent up to 6 days, the kinetics of this induction differed between the two cell types. Despite the in vitro inhibitory effect of genistein, treatment of either HL-205 or K-562-J cells with 150 micrograms/ml genistein for up to 16 h did not alter topoisomerase II activity (as determined by the unknotting assay) in their nuclear extracts. Analysis with the anti-phosphotyrosine PY-20 murine monoclonal antibody indicated that treatment of K-562-J cells with genistein decreased the reactivity of the antibody with two of the cellular proteins. However, no reactivity with the PY-20 antibody was detected in untreated or genistein-treated HL-205 cells. An early event in the HL-205 and K-562-J cells, occurring after only 1 h of treatment with 30-200 micrograms/ml genistein, was the induction of DNA damage as measured by an alkaline elution assay. This damage may be a contributing factor in the genistein-induced cell differentiation in the HL-205 and K-562-J cells.
染料木黄酮是一种拓扑异构酶II和酪氨酸激酶的体外抑制剂，它可抑制人早幼粒白血病HL - 60细胞的克隆群体HL - 205细胞以及人红白血病K - 562细胞的克隆群体K - 562 - J细胞的生长并诱导其分化。成熟的HL - 205细胞获得了粒细胞或单核细胞标志物，即与鼠源OKM1单克隆抗体发生反应、硝基蓝四氮唑染料还原反应的表达以及非特异性酯酶染色。成熟的K - 562 - J细胞用联苯胺染色，这表明存在血红蛋白，即一种红系成熟标志物。尽管HL - 205和K - 562 - J细胞中成熟标志物的获得在长达6天的时间内是时间依赖性的，但两种细胞类型之间这种诱导的动力学有所不同。尽管染料木黄酮具有体外抑制作用，但用150微克/毫升染料木黄酮处理HL - 205或K - 562 - J细胞长达16小时，并未改变其核提取物中的拓扑异构酶II活性（通过解结试验测定）。用抗磷酸酪氨酸PY - 20鼠源单克隆抗体进行分析表明，用染料木黄酮处理K - 562 - J细胞会降低该抗体与两种细胞蛋白的反应性。然而，在未处理或经染料木黄酮处理的HL - 205细胞中未检测到与PY - 20抗体的反应性。在用30 - 200微克/毫升染料木黄酮处理仅1小时后，HL - 205和K - 562 - J细胞中发生的一个早期事件是通过碱性洗脱试验测定的DNA损伤诱导。这种损伤可能是染料木黄酮诱导HL - 205和K - 562 - J细胞分化的一个促成因素。
