Biochemical and Biophysical Research Communications
Up-regulation of PI3K/Akt signaling by 17β-estradiol through activation of estrogen receptor-α, but not estrogen receptor-β, and stimulates cell growth in breast cancer cells
Section snippets
Materials and methods
Materials. Antibodies against phosphatase and tensin homologue deleted on chromosome 10 (PTEN), ERα and ERβ antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against p85, p-Akt (Ser-473), and Akt were purchased from Cell Signaling (Beverly, MD, USA). Fetal calf serum (FBS) and charcoal-dextran treated FBS were obtained from Gibco-BRL (Life Technologies, Grand Island, NY, USA). HBSS (Hanks’ balanced salt solution), 17-β-estradiol, dimethyl sulfoxide
Expression profiling of ER responding to E2 in MCF-7 cells and MDA-MB-231 cells
A previous study demonstrates that uterus shows down-regulation of ERα expression following estrogen exposure of rodents [16]. To determine whether estrogen affects the expression of ER, MCF-7, and MDA-MB-231 cells were treated with E2 (10 nM), and the expression of ER proteins was determined by Western blotting. As shown in Fig. 1A, treatment with E2 resulted in a decrease of ERα level in MCF-7 cells in a time-dependent manner. The ERα protein level at 72 h after E2 treatment was approximately
Acknowledgments
This work was supported by Regional Research Center’s Program of Korean Ministry of Education and Human Resource Development through the Center for Healthcare Technology Development.
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These authors equally contributed to this work.