Correlation of membrane glucocorticoid receptor levels with glucocorticoid-induced apoptotic competence using mutant leukemic and lymphoma cells lines.
We have studied the presence and functional implications of membrane glucocorticoid receptor (mGR) in several wild type (WT) and mutant mouse lymphoid cell lines (nuclear transfer decrease, NT(-); nuclear transfer increase, NT(i); and receptorless, R(-)). Direct fluorescent antibody staining revealed large aggregates of mGR-specific fluorescing antigens in the plasma membrane of the WT and mGR-enriched (mGR(++)) S-49 cells. While R(-) cells totally lacked mGR, this receptor level was low in NT(-) and NT(i) groups. FACS analysis corroborated these results, showing a approximately 4-10-fold difference between the highest mGR levels (mGR(++)) and the R(-) and NT(i) cells. Membrane extracts were analyzed for mGR by immunoblotting. Multiple receptor forms, ranging in M(r) from 94,000 to > 200,000, were observed in the WT cells, while only smaller peptides (85,000-94,000) were found in NT(-) cells. No detectable immunoreactive bands were identified in either membrane or cytosol immunoprecipitates of NT(i) and R(-) cell groups. Within 48 h post dexamethasone exposure > 98% of WT and mGR(++) S-49 cells underwent apoptosis, compared to 0-30% in the mutant cells, albeit the total receptor number is two to three times higher in NT(i) compared to WT. These results suggest a better correlation between the quantity and quality of mGRs (rather than total cellular GRs) and the ability of glucocorticoids (GCs) to lyse lymphoid cells.
Gametchu
Bahiru
B
Department of Pediatrics, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA. gametchu@mcw.edu
Watson
Cheryl S
CS
D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001665:Binding Sites; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient / Q000379:methods; D003907:Dexamethasone / Q000494:pharmacology; D005434:Flow Cytometry; D019085:Fluorescent Antibody Technique, Direct; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D015151:Immunoblotting; D007938:Leukemia / Q000378:metabolism* / Q000473:pathology; D008223:Lymphoma / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D011485:Protein Binding; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D013223:Statistics as Topic; D014407:Tumor Cells, Cultured
D005938:Glucocorticoids; D011965:Receptors, Glucocorticoid; D003907:Dexamethasone