Estrogen Regulates Mitochondrial Morphology through Phosphorylation of Dynamin-related Protein 1 in MCF7 Human Breast Cancer Cells

Abstract

Estrogen affects mitochondrial function in various tissues, but the precise mechanism remains unclear. We, therefore investigated the effect on estrogen-regulated mitochondrial morphology by dynamin-related protein 1 (Drp1) and its Ser616-phosphorylated derivative (pDrp1^Ser616) are involved in mitochondrial fission. MCF7 human breast cancer cells were treated with 17β-estradiol (E₂), an estrogen receptor (ER) α and β antagonist (ICI 182, 780), an ERα antagonist (MPP), and an ERβ antagonist (PHTPP) for 24 hr. The expression of Drp1 and pDrp1^Ser616 was analyzed by western blotting and immunohistochemistry. Mitochondrial morphology was analyzed by transmission electron microscopy (TEM). In control cells, Drp1 was detected in the cytoplasm of all cells while pDrp1 was observed in the cytoplasm of 3.4 ± 1.0% of the total population. After E₂ treatment, pDrp1^Ser616-positive cells comprised 30.6 ± 5.6% of the total population, 10.5 ± 1.7% after E₂ + ICI treatment, 12.4 ± 4.2% after E₂ + MPP treatment, and 24.0 ± 2.2% after E₂ + PHTPP treatment. In ERα knockdown MCF7 cells, pDrp1 expression was decreased after E₂ treatment compared to E₂-treated wild type cells. Tubular pattern mitochondria were found in the control cells but the number of short and small pattern mitochondria (< 0.5 μm²) was significantly increased after E₂ treatment (as observed by TEM). We, therefore concluded that the phosphorylation of Drp1 is important for E₂-dependent mitochondrial morphological changes through ERα.