Abstract
Normal human mammary epithelial cells (HMECs), unlike estrogen receptor-positive (ER+) breast cancers, typically express low nuclear levels of ER (ER-‘poor’). We previously demonstrated that 1.0 μ M tamoxifen (Tam) induced apoptosis in ER-‘poor’ HMECs acutely transduced with human papillomavirus-16 E6 (HMEC-E6) through a rapid mitochondrial signaling pathway. Here, we show that plasma membrane-associated E2-binding sites initiate the rapid apoptotic effects of Tam in HMEC-E6 cells through modulation of AKT activity. At equimolar concentrations, Tam and tamoxifen ethyl bromide (QTam), a membrane impermeant analog of Tam, rapidly induced apoptosis in HMEC-E6 cells associated with an even more rapid decrease in phosphorylation of AKT at serine-473. Treatment of HMEC-E6 cells with 1.0 μ M QTam resulted in a 50% decrease in mitochondrial transmembrane potential, sequential activation of caspase-9 and -3, and a 90% decrease in AKT Ser-473 phosphorylation. The effects of both Tam and QTam were blocked by expression of constitutively active AKT (myristoylated AKT or AKT-Thr308Asp/Ser473Asp). These data indicate that Tam and QTam induce apoptosis in HMEC-E6 cells through a plasma membrane-activated AKT-signaling pathway that results in (1) decreased AKT phosphorylation at Ser-473, (2) mitochondrial membrane depolarization, and (3) activated caspase-9 and -3.
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Acknowledgements
We acknowledge the generous gifts of (1) J Woodgett for the AKT308D/S473D plasmid, (2) D Galloway for the LXSN16E6 retroviral vector containing the HPV-16 E6 coding sequence, and (3) B Katzenellenbogen for the [3H]tamoxifen aziridine. This work is supported by NIH/NCI Grants 2P30CA14236-26 (VLS, ECD), R01CA88799 (VLS), R01CA98441 (VLS), NIH/NIDDK Grant 2P30DK 35816-11 (VLS), DAMD-98-1-851 and DAMD-010919 (VLS), American Cancer Society Award CCE-99898 (VLS), a V-Foundation Award (VLS), a Susan G Komen Breast Cancer Award (VLS, ECD), and a Charlotte Geyer Award (VLS).
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Dietze, E., Troch, M., Bean, G. et al. Tamoxifen and tamoxifen ethyl bromide induce apoptosis in acutely damaged mammary epithelial cells through modulation of AKT activity. Oncogene 23, 3851–3862 (2004). https://doi.org/10.1038/sj.onc.1207480
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DOI: https://doi.org/10.1038/sj.onc.1207480
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