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Estrogen Binding Sites in the Nucleus of Normal and Malignant Human Tissue: Characteristics of the Multiple Nuclear Binding Sites¹

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

The studies presented here describe the effects of the nonionic detergent, Tween 80 and the reducing agent, dithiothreitol (5 mM), on the quantitation of specific nuclear estrogen binding sites, both estrogen receptors (ER) and type II estrogen binding sites (EBS), in human breast cancer. Neither of these agents had any significant effect on the apparent K_d or the amount of ER measured; however, both had a dramatic effect on the amount of type II EBS measured. Two antiestrogens, tamoxifen and nafoxidine, were also tested for their ability to bind to ER and type II EBS in human breast tumors. Both of the antiestrogens were capable of competing with estradiol for ER and type II EBS.

We also present a series of 25 individual human breast cancer specimens which were analyzed for cytoplasmic ER and progesterone receptor by sucrose density gradient centrifugation and for nuclear ER and type II EBS by the sucrose pad assay. Sixty-four % (16 of 25) of the tumors contained nuclear ER and type II EBS when analyzed by the sucrose pad assay. The concentration of type II EBS was strongly correlated to the concentration of cytoplasmic progesterone receptor.

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 64th Annual Meeting of the Endocrine Society, June 16 to 18, 1982, San Francisco, Calif. (20).

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.

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