Glucocorticoid receptor gene mutations in leukemic cells acquired in vitro and in vivo.
Glucocorticoid resistance was investigated in human leukemic CCRF-CEM cells. A mutation (L753F), which renders the human glucocorticoid receptor (hGR) gene functionally hemizygous, was identified in all CEM-derived cell lines analyzed. Allele-specific PCR identified the same mutation in lymph node biopsy material from patient CEM cells. Given the correlation between hGR concentration and glucocorticoid sensitivity, this suggests that loss of functional heterozygosity may result in resistance to glucocorticoid-based chemotherapy. The L753F mutation was probably not responsible for the ontogeny of the disease because it did not appear to be present in all leukemic cells. Thus, it is unlikely that hGR mutations would be detected in leukemic patients at presentation, but they may occur, and be selected for, during treatment. Deletions and point mutations in the hGR gene of cells selected for steroid resistance in vitro were investigated by PCR-single strand conformation polymorphism analysis. Loss of hGR mRNA expression resulted from 5'-deletion of the hGR gene and nonsense mutations in exon 6. These results provide the first evidence for somatic mutation in the hGR gene of a patient with acute lymphoblastic leukemia, offer a potential in vivo mechanism for acquisition of steroid resistance in leukemia, and suggest that screening for additional in vivo mutations will require analysis of genomic DNA.
Hillmann
A G
AG
Department of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799, USA.
Ramdas
J
J
Multanen
K
K
Norman
M R
MR
Harmon
J M
JM
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D015252:Deoxyribonucleases, Type II Site-Specific; D006801:Humans; D007938:Leukemia / Q000235:genetics* / Q000473:pathology; D008198:Lymph Nodes / Q000473:pathology; D009154:Mutation; D017354:Point Mutation; D012150:Polymorphism, Restriction Fragment Length; D018807:Polymorphism, Single-Stranded Conformational; D011965:Receptors, Glucocorticoid / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes; D014407:Tumor Cells, Cultured
D011965:Receptors, Glucocorticoid; C024669:endodeoxyribonuclease AluI; C041666:endodeoxyribonuclease BclI; D015252:Deoxyribonucleases, Type II Site-Specific