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Estrogen Binding Sites in the Nucleus of Normal and Malignant Human Tissue: Optimization of an Exchange Assay for the Measurement of Specific Binding¹

Departments of Pathology [J. S. S., W. B. P.] and Cell Biology [J. S. S., B. M. M., J. H. C., W. B. P.], Baylor College of Medicine, Houston, Texas 77030

An exchange assay has been validated and used to measure the total concentration of estrogen receptors in the nuclei of cells from human breast tumors and myometrium. Tissue homogenates were centrifuged through 1.2 M sucrose pads to separate crude chromatin from soluble and membranous fractions. The yield of DNA by this procedure was approximately 80%. Total binding sites were measured by incubating the sedimented pellets with tritiated estradiol at 30° for 60 min and then precipitating the receptor-steroid complexes with protamine sulfate. Saturation analysis by this procedure provides evidence for the presence of a specific, nuclear estrogen-binding site in addition to the established estrogen receptor in both these tissues.

¹ This work was supported by Grants CA-25586 and CA-26452 from the National Cancer Institute, NIH, USPHS, Department of Health and Human Services. A preliminary report of these data was presented at the 73rd Annual Meeting of the American Association for Cancer Research, April 28 to May 1, 1982, St. Louis, Mo. (25).

² To whom requests for reprints should be addressed, at Department of Pathology, Baylor College of Medicine, Houston, Texas 77030.

Received 1/13/82. Accepted 8/ 3/82.