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. 1987 Dec;7(12):4211–4217. doi: 10.1128/mcb.7.12.4211

Glucocorticoid-resistant lymphoma cell variants that contain functional glucocorticoid receptors.

S K Rabindran 1, M Danielsen 1, M R Stallcup 1
PMCID: PMC368102  PMID: 2830485

Abstract

A mouse T-lymphosarcoma cell line stably infected with mouse mammary tumor virus (MMTV) was used as the parent line for a genetic analysis of two glucocorticoid hormone responses, hormone-induced cytolysis and stimulation of viral gene expression. Variants were selected for survival and elevated expression of MMTV proteins in the presence of the steroid. The MMTV marker provided a sensitive test for glucocorticoid receptor (GR) function in the hormone-resistant variants. This strategy resulted in the isolation of two novel types of hormone-resistant variants. One type of variant with only about 25% of the level of GR found in the parent line was resistant to the cytolytic effects of glucocorticoid but produced increased levels of MMTV gene products in response to the hormone. This variant phenotype demonstrated that the MMTV response requires fewer GR than the cytolytic response. Another variant, which required approximately 100-fold higher concentrations of hormone than the wild-type cells for both responses, apparently contained GR with altered hormone-binding properties.

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Selected References

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  1. Bourgeois S., Newby R. F. Correlation between glucocorticoid receptor and cytolytic response of murine lymphoid cell lines. Cancer Res. 1979 Nov;39(11):4749–4751. [PubMed] [Google Scholar]
  2. Bourgeois S., Newby R. F. Diploid and haploid states of the glucocorticoid receptor gene of mouse lymphoid cell lines. Cell. 1977 Jun;11(2):423–430. doi: 10.1016/0092-8674(77)90060-5. [DOI] [PubMed] [Google Scholar]
  3. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  4. Danielsen M., Northrop J. P., Ringold G. M. The mouse glucocorticoid receptor: mapping of functional domains by cloning, sequencing and expression of wild-type and mutant receptor proteins. EMBO J. 1986 Oct;5(10):2513–2522. doi: 10.1002/j.1460-2075.1986.tb04529.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Danielsen M., Peterson D. O., Stallcup M. R. Immunological selection of variant mouse lymphoid cells with altered glucocorticoid responsiveness. Mol Cell Biol. 1983 Jul;3(7):1310–1316. doi: 10.1128/mcb.3.7.1310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Danielsen M., Stallcup M. R. Down-regulation of glucocorticoid receptors in mouse lymphoma cell variants. Mol Cell Biol. 1984 Mar;4(3):449–453. doi: 10.1128/mcb.4.3.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gehring U., Mugele K., Ulrich J. Cellular receptor levels and glucocorticoid responsiveness of lymphoma cells. Mol Cell Endocrinol. 1984 Jun;36(1-2):107–113. doi: 10.1016/0303-7207(84)90089-3. [DOI] [PubMed] [Google Scholar]
  8. Godowski P. J., Rusconi S., Miesfeld R., Yamamoto K. R. Glucocorticoid receptor mutants that are constitutive activators of transcriptional enhancement. Nature. 1987 Jan 22;325(6102):365–368. doi: 10.1038/325365a0. [DOI] [PubMed] [Google Scholar]
  9. Grove J. R., Dieckmann B. S., Schroer T. A., Ringold G. M. Isolation of glucocorticoid-unresponsive rat hepatoma cells by fluorescence-activated cell sorting. Cell. 1980 Aug;21(1):47–56. doi: 10.1016/0092-8674(80)90113-0. [DOI] [PubMed] [Google Scholar]
  10. Harmon J. M., Schmidt T. J., Thompson E. B. Molybdate-sensitive and molybdate-resistant activation-labile glucocorticoid-receptor mutants of the human lymphoid cell line CEM-C7. J Steroid Biochem. 1984 Sep;21(3):227–236. doi: 10.1016/0022-4731(84)90274-7. [DOI] [PubMed] [Google Scholar]
  11. Harmon J. M., Thompson E. B. Isolation and characterization of dexamethasone-resistant mutants from human lymphoid cell line CEM-C7. Mol Cell Biol. 1981 Jun;1(6):512–521. doi: 10.1128/mcb.1.6.512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harris A. W., Bankhurst A. D., Mason S., Warner N. L. Differentiated functions expressed by cultured mouse lymphoma cells. II. Theta antigen, surface immunoglobulin and a receptor for antibody on cells of a thymoma cell line. J Immunol. 1973 Feb;110(2):431–438. [PubMed] [Google Scholar]
  13. Hollenberg S. M., Giguere V., Segui P., Evans R. M. Colocalization of DNA-binding and transcriptional activation functions in the human glucocorticoid receptor. Cell. 1987 Apr 10;49(1):39–46. doi: 10.1016/0092-8674(87)90753-7. [DOI] [PubMed] [Google Scholar]
  14. Horibata K., Harris A. W. Mouse myelomas and lymphomas in culture. Exp Cell Res. 1970 Apr;60(1):61–77. doi: 10.1016/0014-4827(70)90489-1. [DOI] [PubMed] [Google Scholar]
  15. Johnson D. M., Newby R. F., Bourgeois S. Membrane permeability as a determinant of dexamethasone resistance in murine thymoma cells. Cancer Res. 1984 Jun;44(6):2435–2440. [PubMed] [Google Scholar]
  16. Norman M. R., Thompson E. B. Characterization of a glucocorticoid-sensitive human lymphoid cell line. Cancer Res. 1977 Oct;37(10):3785–3791. [PubMed] [Google Scholar]
  17. Northrop J. P., Danielsen M., Ringold G. M. Analysis of glucocorticoid unresponsive cell variants using a mouse glucocorticoid receptor complementary DNA clone. J Biol Chem. 1986 Aug 25;261(24):11064–11070. [PubMed] [Google Scholar]
  18. Northrop J. P., Gametchu B., Harrison R. W., Ringold G. M. Characterization of wild type and mutant glucocorticoid receptors from rat hepatoma and mouse lymphoma cells. J Biol Chem. 1985 May 25;260(10):6398–6403. [PubMed] [Google Scholar]
  19. Pfahl M., Kelleher R. J., Jr, Bourgeois S. General features of steroid resistance on lymphoid cell lines. Mol Cell Endocrinol. 1978 Apr;10(2):193–207. doi: 10.1016/0303-7207(78)90125-9. [DOI] [PubMed] [Google Scholar]
  20. Rabindran S. K., Danielsen M., Firestone G. L., Stallcup M. R. Glucocorticoid-dependent maturation of viral proteins in mouse lymphoma cells: isolation of defective and hormone-independent cell variants. Somat Cell Mol Genet. 1987 Mar;13(2):131–143. doi: 10.1007/BF01534693. [DOI] [PubMed] [Google Scholar]
  21. Rabindran S. K., Stallcup M. R. Mouse lymphoma cell variants with genetically dominant alterations in the maturation of viral glycoproteins. Mol Endocrinol. 1987 Jul;1(7):491–499. doi: 10.1210/mend-1-7-491. [DOI] [PubMed] [Google Scholar]
  22. Sibley C. H., Tomkins G. M. Isolation of lymphoma cell variants resistant to killing by glucocorticoids. Cell. 1974 Aug;2(4):213–220. doi: 10.1016/0092-8674(74)90013-0. [DOI] [PubMed] [Google Scholar]
  23. Sibley C. H., Tomkins G. M. Mechanisms of steroid resistance. Cell. 1974 Aug;2(4):221–227. doi: 10.1016/0092-8674(74)90014-2. [DOI] [PubMed] [Google Scholar]
  24. Simons S. S., Jr, Pumphrey J. G., Rudikoff S., Eisen H. J. Identification of cysteine 656 as the amino acid of hepatoma tissue culture cell glucocorticoid receptors that is covalently labeled by dexamethasone 21-mesylate. J Biol Chem. 1987 Jul 15;262(20):9676–9680. [PubMed] [Google Scholar]
  25. Simons S. S., Jr, Schleenbaker R. E., Eisen H. J. Activation of covalent affinity labeled glucocorticoid receptor-steroid complexes. J Biol Chem. 1983 Feb 25;258(4):2229–2238. [PubMed] [Google Scholar]
  26. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Vanderbilt J. N., Miesfeld R., Maler B. A., Yamamoto K. R. Intracellular receptor concentration limits glucocorticoid-dependent enhancer activity. Mol Endocrinol. 1987 Jan;1(1):68–74. doi: 10.1210/mend-1-1-68. [DOI] [PubMed] [Google Scholar]
  28. Westphal H. M., Mugele K., Beato M., Gehring U. Immunochemical characterization of wild-type and variant glucocorticoid receptors by monoclonal antibodies. EMBO J. 1984 Jul;3(7):1493–1498. doi: 10.1002/j.1460-2075.1984.tb02001.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Yamamoto K. R., Gehring U., Stampfer M. R., Sibley C. H. Genetic approaches to steroid hormone action. Recent Prog Horm Res. 1976;32:3–32. doi: 10.1016/b978-0-12-571132-6.50008-7. [DOI] [PubMed] [Google Scholar]

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