Identification of a glucocorticoid receptor in the human leukemia cell line K562.
The binding of [3H]dexamethasone to the cytosol fraction prepared from the human leukemia cell line K562 was studied with a competitive binding assay. Specific, saturable binding was identified by incubating cytosol with increasing concentrations of [3H]dexamethasone in the presence and absence of nonlabeled dexamethasone. A Scatchard plot of the data was linear, suggesting the presence of a single class of binding sites with KD = 2.49 +/- 0.23 x 10(-8)M. The binding sites appear to be protein in nature, since specific binding was reduced by treatment of the cytosol with trypsin, pronase, and heat; neither DNase nor RNase affected the binding. Binding was also reduced in the absence of alpha-thioglycerol and in the presence of p-chloromercuribenzoate and N-ethylamaleimide, suggesting that optimal binding activity requires reduced sulfhydryl groups. The binding site appears to be specific for glucocorticoids as evaluated in competition studies. Finally, glucocorticoids were found to inhibit the clonal growth of K562 cells in vitro, suggesting a potential role for glucocorticoid binding sites in the modulation of K562 cell proliferation.
D000818:Animals; D001667:Binding, Competitive; D002460:Cell Line; D003114:Colony-Forming Units Assay; D003600:Cytosol / Q000378:metabolism; D003907:Dexamethasone / Q000378:metabolism; D006801:Humans; D007942:Leukemia, Experimental / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000032:analysis*; D011987:Receptors, Steroid / Q000032:analysis*
D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D003907:Dexamethasone