Membrane Estrogen Signaling Enhances Tumorigenesis and Metastatic Potential of Breast Cancer Cells via Estrogen Receptor-alpha 36 (ER alpha 36)

• Type of Material: Article
• Authors: Barbara Boyan, Emory University; Reyhaan A Chaudhri, Emory University; Rene Olivares-Navarrete, Georgia Institute of Technology; Natalia Cuenca, Georgia Institute of Technology; Agreen Hadadi, Georgia Institute of Technology; Zvi Schwartz, Georgia Institute of Technology
• Language: English
• Date: 2012-03-02
• Publisher: American Society for Biochemistry and Molecular Biology
• Publication Version: Final Published Version
• Copyright Statement: © 2012 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.
• Final Published Version (URL): http://dx.doi.org/10.1074/jbc.M111.292946
• Title of Journal or Parent Work: Journal of Biological Chemistry
• Volume: 287
• Issue: 10
• Start Page: 7169
• End Page: 7181
• Grant/Funding Information: TL1 RR025010 (Clinical and Translational Science Award Program) from NCRR; Price Gilbert, Jr., Foundation; National Institutes of Health Grants UL1 RR025008 from USPHS
• Keywords: GROWTH-PLATE CHONDROCYTES; EXPRESSION; ER-ALPHA; PLASMA-MEMBRANE; ACTIVATION; Science & Technology; PROTEIN-KINASE-C; Biochemistry & Molecular Biology; VARIANT; EPITHELIAL-MESENCHYMAL TRANSITION; RECEPTOR-ALPHA; Life Sciences & Biomedicine; BONE METASTASIS

Abstract

Protein kinase C (PKC) signaling can be activated rapidly by 17β-estradiol (E 2) via nontraditional signaling in ERα- positive MCF7 and ERα-negative HCC38 breast cancer cells and is associated with tumorigenicity. Additionally, E 2 has been shown to elicit anti-apoptotic effects in cancer cells counteracting pro-apoptotic effects of chemotherapeutics. Supporting evidence suggests the existence of a membrane-associated ER that differs from the traditional receptors, ERαand ERβ. Our aim was to identify the ER responsible for rapid PKC activation and to evaluate downstream effects, such as proliferation, apoptosis, and metastasis. RT-PCR, Western blot, and immunofluorescence were used to determine the presence of ER splice variants in multiple cell lines. E 2 effects on PKC activity were measured with and without ER-blocking antibodies. Cell proliferation was determined by [ 3H]thymidine incorporation, and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, (MTT) whereas apoptosis was determined by DNA fragmentation and TUNEL. Quantitative RT-PCR and sandwich ELISA were used to determine the effects on metastatic factors. The role of membrane-dependent signaling in cancer cell invasiveness was examined using an in vitro assay. The results indicate the presence of an ERα splice variant, ERα36, in ERα-positive MCF7 and ERα-negative HCC38 breast cancer cells, which localized to plasma membranes and rapidly activated PKC in response to E 2, leading to deleterious effects such as enhancement of proliferation, protection against apoptosis, and enhancement of metastatic factors. These findings propose ERα36 as a novel target for the development of therapies that can prevent progression of breast cancer in the primary tumor as well as during metastasis. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.