Analysis of activated and nonactivated cytoplasmic glucocorticoid-receptor complexes from human leukemia cells by rapid DNA-diethylaminoethyl minicolumn chromatography.
Glucocorticoid-receptor complexes in cytoplasm from normal lymphoid cells incubated with [3H]dexamethasone can be resolved into three different components. Two of these correspond to the well-established activated and nonactivated forms, while the third appears similar to the mero-receptor complex first described by Sherman et al. (Fed. Proc., 37: 167-173, 1978). Based on their differential affinities for DNA- and DEAE-cellulose in buffers of low ionic strength (the activated complex binds to DNA- and DEAE-cellulose; the nonactivated complex binds to only DEAE-cellulose; the mero-receptor-like complex binds to neither), we have developed a rapid minicolumn chromatographic procedure for separating these forms, and have applied it to examine the relative proportion of different complexes in cytosols from cells of leukemia patients. All samples from nine patients with chronic lymphocytic leukemia contained these three complexes in proportions similar to those seen with normal lymphoid tissue. Cytosols from six of eight specimens from patients with acute nonlymphocytic leukemia contained a lower proportion of activated complexes and a higher proportion of mero-receptor-like complexes than cytosols from normal or chronic lymphocytic leukemia cells. Whether such differences in the properties of cytosolic complexes are due to degradation taking place after the cells are broken, and whether they can be correlated to in vivo therapeutic response, is not known, but studies are in progress to answer these questions. The minicolumn procedure described here offers a simple and reliable method for these purposes.
D000293:Adolescent; D000328:Adult; D000368:Aged; D002846:Chromatography, Affinity / Q000379:methods; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D003600:Cytosol / Q000378:metabolism; D003636:DEAE-Cellulose; D004247:DNA; D003907:Dexamethasone / Q000378:metabolism*; D005260:Female; D006801:Humans; D007202:Indicators and Reagents; D007700:Kinetics; D007938:Leukemia / Q000378:metabolism*; D007945:Leukemia, Lymphoid / Q000378:metabolism; D008214:Lymphocytes / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D011965:Receptors, Glucocorticoid / Q000302:isolation & purification / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*
D007202:Indicators and Reagents; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D003907:Dexamethasone; D004247:DNA; D003636:DEAE-Cellulose