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Effect of Adrenal Manipulation on Glucocorticoid Receptors in MM1 Hamster Melanoma¹

Department of Pharmacology [L. R. S., C. W. B.] and Division of Surgical Oncology [L. R. S., C. W. B.], University of Illinois; Cook County [W. F. L.] and West Side Veterans Administration Hospitals; and the Hektoen Institute for Medical Research, Chicago, Illinois 60680

Cytosol glucocorticoid receptor of hamster Malignant Melanoma No. 1 (MM1) was characterized by dextran-coated charcoal and sucrose gradient assays. MM1 contains a specific, saturable, high-affinity (K_d 2.5 nM; 73.3 fmol/mg cytosol protein) receptor for glucocorticoid. The receptor sedimented at 7 to 8S in low-salt buffer and 5.5S in 0.4 M KCI and was sensitive to proteolysis by trypsin. Glucocorticoid receptor was inactivated by 40% (NH₄)₂SO₄. Addition of 20 mM molybdate to the homogenizing buffer prevented inactivation. Adrenalectomy increased MM1 receptor content without altering affinity. Chronic exposure of intact or adrenalectomized hamsters to gluco- and high-dose mineralocorticoid significantly decreased the amount of unbound glucocorticoid receptors available for binding. Exposure of intact hamsters to high doses of mineralocorticoid also decreased apparent receptor affinity.

These results suggest that hamster MM1 contains a high-affinity cytosol receptor for glucocorticoids similar to that of other glucocorticoid-responsive tissues, which may mediate the action of adrenal steroids on tumor behavior.

¹ Supported by the University of Illinois Campus Research Board, Grant 1661; the Cancer Research Fund; and the Carol Thomas Brigham Fund. This is Paper 2, Biological Behavior of Hamster MM1, of a series.

² Submitted as partial fulfillment of requirements for Doctor of Philosophy, Department of Pharmacology, University of Illinois Medical Center, Chicago, Ill. Present address: Department of Pediatrics, University of Utah Medical Center, Salt Lake City, Utah 84132.

³ To whom requests for reprints should be addressed, at Division of Surgical Oncology, University of Illinois Medical Center, 840 S. Wood St., Chicago, Ill. 60612.

Received 10/13/81. Accepted 3/11/82.

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