A glucocorticoid-resistant rat hepatoma cell variant contains functional glucocorticoid receptor.
The mechanism of glucocorticoid resistance was studied in a rat hepatoma cell variant (6.10.2) which contains low levels of glucocorticoid receptor. These cells seem to have lost glucocorticoid-induced transcriptional responses as measured by the induction of expression of stably integrated mouse mammary tumor virus gene and the endogenous tyrosine aminotransferase gene, as well as the transcriptional suppression of glucocorticoid receptor gene expression. However, characterization of the glucocorticoid resistance in 6.10.2 cells revealed that the receptor is indistinguishable from the wild-type receptor with respect to hormone binding and affinity for both nonspecific and specific DNA sequences. The levels of the receptor mRNA and the total immunoreactive protein found in 6.10.2 cells were about 20% of those found in wild-type cells. Further analysis of 6.10.2 cells demonstrated that the receptor was indeed biologically functional. First, treatment of 6.10.2 cells with 8-bromo-cAMP elevated the endogenous glucocorticoid receptor levels 2-fold and restored responsiveness to glucocorticoids. Second, pretreatment of the cells with cycloheximide also led to acquisition of cellular responsiveness to glucocorticoids. We propose that there exists a "threshold" level of glucocorticoid receptor which is required for responsiveness and that under normal culture conditions, the level of glucocorticoid receptor in 6.10.2 cells is below this threshold. However, glucocorticoid responsiveness can be restored by raising the glucocorticoid receptor level above the threshold with 8-bromo-cAMP or, alternatively, by removing the threshold barrier with cycloheximide.
Dong
Y
Y
Department of Medical Nutrition, Karolinska Institute, Huddinge University Hospital, Sweden.
Cairns
W
W
Okret
S
S
Gustafsson
J A
JA
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002472:Cell Transformation, Viral; D002846:Chromatography, Affinity; D003600:Cytosol / Q000378:metabolism; D004273:DNA, Neoplasm / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology*; D004351:Drug Resistance; D014644:Genetic Variation; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D014444:Tyrosine Transaminase / Q000378:metabolism
D004273:DNA, Neoplasm; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid; D003907:Dexamethasone; D014444:Tyrosine Transaminase