Summary
We have used in vitro DNA binding assays as a measure of estrogen receptor (ER) function in human breast tumors. We found that the majority of ER+ (25 ER+/progesterone receptor [PgR]+, and 25 ER+/PgR−) tumors we examined were capable of binding consensus estrogen response element (ERE) oligonucleotides in this assay system. We found significant proteolytic activity in many of the tumors such that protease inhibitors were found to be essential during the preparation of tumor extracts. We next applied direct sequence analysis of the ER DNA binding domain of several of these tumors, and determined that the ER+/PgR− breast tumors did not contain mutations within the DNA binding domain which might explain their apparent discordant receptor phenotype. We did identify an alternatively spliced ER variant missing exon 3 of the DNA binding domain. This variant was unable to function as a transcriptional inducer of an estrogen-responsive reporter in a yeast assay system. Furthermore, the exon 3 ER deletion variant was expressed at equivalent levels in all of the ER+ breast tumors, so that it does not appear to be involved in the evolution of the ER+/PgR− breast cancer phenotype.
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William L. McGuire was a Clinical Professor of the American Cancer Society until his untimely death on March 25, 1992, while this work was in progress. This report is dedicated to his memory, with gratitude for his leadership and his support.
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Fuqua, S.A.W., Allred, D.C., Elledge, R.M. et al. The ER-positive / PgR-negative breast cancer phenotype is not associated with mutations within the DNA binding domain. Breast Cancer Res Tr 26, 191–202 (1993). https://doi.org/10.1007/BF00689692
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DOI: https://doi.org/10.1007/BF00689692