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Tamoxifen aziridine binding to cytosolic proteins from human breast specimens is negatively associated with estrogen receptors, progesterone receptors, pS2, and cathepsin-D

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Abstract

[3H]Tamoxifen Aziridine ([3H]TAZ) is a derivative of the antiestrogen tamoxifen that covalently labels the Estrogen Receptor (ER), and perhaps other uncharacterized proteins. In a previous article we described that [3H]TAZ binds to a cytosolic protein from human uterine tissues that shares some, but not all, the ER properties. Here we have extended these studies to [3H]TAZ binding to cytosol proteins from human breast cancer specimens, and studied its quantitative association with other molecular markers and clinico-pathological variables. Cytosols were obtained in hypotonic buffer containing 20 mM molybdate and protease inhibitors, incubated with [3H]TAZ, and subjected to Sucrose Gradient Analysis (SGA). A [3H]TAZ labeled peak that consistently migrated with the 4S fractions was found in most of the assayed cytosols (range of 0 to 1278 fmol/mg p.). The 4S peak of [3H]TAZ was partially inhibited by both estrogens and antiestrogens. When [3H]E2 was used instead of [3H]TAZ, only an 8S peak was detected. [3H]TAZ was covalently bound to a protein with an apparent MW of 65 kDa, as determined by SDS-PAGE and fluorography. The mean of [3H]TAZ binding was significantly higher in the subgroups of samples classified as ER-, PR-, pS2- or cathepsin D-, than in the respective positive subgroups (P < 0.01 in all the cases). [3H]TAZ binding was not associated with clinico-pathological variables, except that its mean was significantly larger in tumors larger than 5 cm than in smaller tumors. These results, and those previously reported, suggest that: 1) [3H]TAZ labels a cytosolic protein present in human breast cancers and uterine tissues that does not share all the ER properties, and 2) the [3H]TAZ binding by breast cancer cytosols is negatively associated with markers of estrogenic dependency, and its quantification may provide valuable information on antiestrogen responsiveness of a given tumor.

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Authors and Affiliations

  1. Laboratorio de Fisiología, Dept. Endocrinología Celular y Molecular, Centro de Ciencias de la Salud, Universidad de Las Palmas de Gran Canaria, Spain

    Domingo Navarro, Hilario Doreste, Juan C. Díaz-Chico & Bonifacio N. Díaz-Chico

  2. Servicio de Anatomía Patológica, Hospital Insular de Gran Canaria, Spain

    Juan J. Cabrera

  3. Servicio de Oncología Médica, Hospital de La Candelaria, Santa Cruz de Tenerife, Islas Canarias, Spain

    Manuel Morales

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  1. Domingo Navarro
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  5. Juan C. Díaz-Chico
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  6. Bonifacio N. Díaz-Chico
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Navarro, D., Doreste, H., Cabrera, J.J. et al. Tamoxifen aziridine binding to cytosolic proteins from human breast specimens is negatively associated with estrogen receptors, progesterone receptors, pS2, and cathepsin-D. Breast Cancer Res Treat 50, 155–166 (1998). https://doi.org/10.1023/A:1006062510883

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