Human glucocorticoid receptor gene deletion following exposure to cancer chemotherapeutic drugs and chemical mutagens.
The sensitivity of the human glucocorticoid receptor (hGR) gene to mutagenesis by the cancer chemotherapeutic drugs adriamycin, bleomycin, and chlorambucil was evaluated using glucocorticoid-sensitive (dexs) subclones of the human leukemic cell line CEM-C7. Treatment of cells with either bleomycin or chlorambucil increased the frequency of glucocorticoid-resistant (dexr) clones 3.3- and 10-fold, respectively. Measurement of steroid-binding activity in intact dexr cells demonstrated that the predominant phenotype of drug-induced dexr clones was receptorless (r-). dexs CEM cells express only one functional hGR allele and, in addition, are heterozygous for a BclI restriction fragment length polymorphism in the hGR gene (L. A. Palmer and J. M. Harmon, Cancer Res., 51:5224-5231, 1991). To determine the basis of the r- phenotype, BclI digests of genomic DNA isolated from r+ and r- cell lines were examined for the presence of the polymorphic 2.4- and 4.4-kilobase digestion products. A deletion of all or part of the hGR gene was demonstrated by the absence of the 4.4-kilobase fragment in one of two bleomycin-induced dexr clones, as well as the ICR191-induced dexr cell line ICR27TK.3. Cytogenetic analysis of ICR27TK.3 showed that the distal portion of the long arm of one chromosome 5 had been replaced with a portion of chromosome 15. Thus, in at least two dexr cell lines, deletions and/or chromosome breaks in the hGR locus appear to account for the r- phenotype.
Palmer
L A
LA
Department of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799.
Hukku
B
B
Harmon
J M
JM
D000483:Alleles; D001761:Bleomycin / Q000494:pharmacology*; D002478:Cells, Cultured; D002699:Chlorambucil / Q000494:pharmacology*; D002895:Chromosomes, Human, Pair 5; D003907:Dexamethasone / Q000378:metabolism; D004317:Doxorubicin / Q000494:pharmacology*; D017353:Gene Deletion; D006801:Humans; D007621:Karyotyping; D010641:Phenotype; D012333:RNA, Messenger / Q000032:analysis*; D011965:Receptors, Glucocorticoid / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism
D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid; D001761:Bleomycin; D002699:Chlorambucil; D003907:Dexamethasone; D004317:Doxorubicin