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Comparison of Dextran-coated Charcoal and Sucrose Density Gradient Analyses of Estrogen and Progesterone Receptors in Human Breast Cancer¹

Oncology Laboratory, Section of Medical Oncology, Department of Medicine and Department of Biochemistry [K. M. A., P. B., M. M., C. H.], and Department of Surgery [S. E.], Rush Medical College, Chicago, Illinois 60612

Results of estrogen receptor (ER) determinations in 300 primary breast cancers and of progesterone receptor (PR) in the last 100 specimens are reported. In the first 100 samples, a single-point dextran-coated charcoal (DCC) assay and sucrose density gradient (SDG) centrifugation were compared with agreement in 37 ER-positive and 58 ER-negative samples. Material from three additional patients was positive (>250 fmol/g wet weight) by DCC and negative (<3 fmol/g wet weight) by SDG centrifugation analysis. Two other patients positive by gradient were "borderline" by DCC analysis.

For the next 200 samples, results of the single-point DCC assay were used to select patients for SDG centrifugation, and all borderline (>3 but <10 fmol/mg protein) or positive (10 fmol/mg protein) samples were subjected to SDG centrifugation. Agreement between the two methods was satisfactory. The last 100 cytosols were also examined for PR by both procedures. Fifty-three % of these 200 patients were ER positive. Forty-two % of the last 100 samples were ER positive and PR positive, 33% were ER negative and PR negative, 12% were ER positive and PR negative, and 13% were ER negative and PR positive. Overall, 54 and 55% were ER positive and PR positive, respectively. If 3 fmol/mg protein were considered the lower limit for positive samples, 56 and 63% were ER positive or PR positive.

Major differences between the results of DCC and SDG analysis of PR were found with values of the latter usually much less than those of the former. Inclusion of 20 mM sodium molybdate during labeling with steroid reduced these discrepancies. In some samples, samller increases in the amount of ER were obtained.

Although ER and PR values tended to be positively correlated, they could also occur apparently independently of each other. About one-half of the ER-negative and PR-positive samples and one-half of the ER-positive and PR-negative samples were from premenopausal patients. The number of ER-positive and PR-positive samples increased with the age of the patients; this was not true of the ER-positive and PR-negative or the ER-negative and PR-positive categories. The highest receptor values were obtained with samples from patients 50 years or older. ER and PR in buffer of low ionic strength were composed chiefly of 8S receptors; for PR, this was particularly true in the presence of sodium molybdate. Of 100 metastatic samples analyzed for ER and 30 for both receptors, the overall distribution resembled that of primary tumors.

The use of sodium molybdate increased the percentage of PR-positive samples and the absolute amounts of receptor measured. Effects of this salt on ER assays were less evident although, in some samples, smaller increases in the amount of ER detected occurred. This salt should be useful for studying certain aspects of steroid receptor "processing" from 8S to other receptor moieties.

¹ This work was supported in part by the National Cancer Institute Grant CA-22246, the Illinois Division of the American Cancer Society Grant 77-62, the Wadsworth Memorial Fund, and the Charles Perlia Memorial Fund.

² To whom requests for reprints should be addressed.

Received 9/ 4/79. Accepted 8/ 6/80.

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