Induction of differentiation and DNA strand breakage in human HL-60 and K-562 leukemia cells by genistein.
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.
Constantinou
A
A
Biological and Medical Research Division, Argonne National Laboratory, IL 60439.
Kiguchi
K
K
Huberman
E
E
D002454:Cell Differentiation / Q000187:drug effects; D004247:DNA / Q000187:drug effects* / Q000378:metabolism; D004249:DNA Damage; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology*; D019833:Genistein; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology*; D007938:Leukemia / Q000473:pathology*; D008970:Molecular Weight; D010766:Phosphorylation; D059005:Topoisomerase II Inhibitors; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism
D005419:Flavonoids; D007529:Isoflavones; D059005:Topoisomerase II Inhibitors; D014443:Tyrosine; D004247:DNA; D019833:Genistein; D004250:DNA Topoisomerases, Type II