The effects of ras gene expression on glucocorticoid receptors in mouse fibroblasts.
Analysis of induction of glutamine synthetase activity by dexamethasone showed a 2-fold increase in NIH3T3 but no change in NIH3T3 ras (EJ-ras) cells. The observed increase could be abolished by the antagonist RU486. The lack of response in ras transformed cells might reflect oncoprotein effects on the glucocorticoid receptor (GR). Several GR parameters were studied in order to clarify this point. Total GR level was the same for both cells; cytoplasmic receptor level however, was 3 times lower in NIH3T3 ras than in NIH3T3 cells. Hormone-receptor binding affinity, specificity, thermostability, sedimentation coefficient, molecular weight as well as the cytoplasmic GR transformation ratio were similar for the two cell lines. On the other hand, the fraction of the total receptor pool involved with the recycling process was approximately 20% lower in NIH3T3 ras than in NIH3T3 cells. After 24 h of dexamethasone treatment, no GR down regulation was observed in NIH3T3 ras cells, whereas normal NIH3T3 cells exhibited a decrease of GR binding capacity around 80%. Further studies are necessary to define the mechanisms underlying the association between glucocorticoid insensitivity, and modifications in the GR nuclear/cytoplasmic ratio, in the recycling GR fraction and in the down-regulation process observed in ras transformed cells.
Martins
V R
VR
Departamento de Clinica Medica, Faculdade de Medicina, Universidade de São Paulo, Brasil.
Brentani
M M
MM
D000818:Animals; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D002482:Cellulose / Q000031:analogs & derivatives; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D003600:Cytosol / Q000378:metabolism; D004247:DNA; D003907:Dexamethasone / Q000378:metabolism / Q000494:pharmacology*; D011905:Genes, ras; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D015735:Mifepristone / Q000494:pharmacology; D011965:Receptors, Glucocorticoid / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*
C033190:DNA-cellulose; D011965:Receptors, Glucocorticoid; D015735:Mifepristone; D003907:Dexamethasone; D002482:Cellulose; D004247:DNA; D005974:Glutamate-Ammonia Ligase