Modulatory Effect of Nonesterified Fatty Acids on Structure and Binding Characteristics of Estrogen Receptor from MCF-7 Human Breast Cancer Cells

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Abstract
Arachidonic, docosahexaenoic and oleic acids were found to produce a dose-dependent cytotoxic effect on MCF-7 cells; palmitic and stearic acids were totally ineffective in this regard suggesting that solely unsaturated fatty acids were able to arrest mammary tumor cell growth. Similarly, only former acids were able to decrease the binding capacity and affinity (increase Kd value) of the cells for 3H-E2 in a dose-dependent manner. In the case of arachidonic acid (the reference fatty acid), this decrease was associated with a slight cleavage of the native 67 KDa estrogen receptor (ER) into 50 and 25–30 KDa peptides as demonstred by sequential labeling of high-salt cell extracts with 3H-tamoxifen aziridine, specific immunoadsorption with H-222 anti-ER monoclonal antibody, SDS-PAGE and fluorography. Both, modifications in binding characteristics of ER and cleavage of the native 67 KDa receptor were found to be extremely marked when unsaturated fatty acids were directly added to the high-salt cell extracts. This clear influence on the ER structure was reflected on enzyme immunological assay (EIA) by a reduction of ER immunoreactivity of approximatly 50% in presence of arachidonic acid. Our observations are discussed in terms of possible interference of unsaturated fatty acids either through transmembrane modulation of phosphokinases and/or phospholipases implicated in ER mechanism of action, or through an intracellular interaction between ER and these acids acting as second messengers in regulation of cellular functions.