Citation

  • Authors: Mohammadi Ghahhari N. et al.
  • Year: 2022
  • Journal: Nat Commun 13 2104
  • Applications: in vitro / DNA / jetOPTIMUS
  • Cell types:
    1. Name: MCF-7
    2. Name: T-47D
      Description: Human breast ductal carcinoma cells
      Known as: T 47

Method

24 h prior to transfections, 4 × 104 cells for MCF7, MCF7-V, MCF7-V-ZEB1, or T-47D cells were seeded in the complete or steroid-deprived medium in each well of a 24-well plate. Cells were transiently transfected with the indicated plasmids and corresponding firefly luciferase reporters and pRL-CMV for renilla luciferase expression using jetOPTIMUS. After 8 h, the medium was changed and 24 h after transfection specific treatments were added for 18–24 h including vehicle, E2 (10 nM), FI (10 μM forskolin + 100 μM IBMX), P4 (10 nM), and TGFβ1 (10 ng/ml). Luciferase activity was measured with the dual-luciferase reporter assay (Promega). The firefly and renilla luciferase activities were measured with a bioluminescence plate reader. Normalization to the renilla luciferase internal control was performed to quantify the activity.

Abstract

The epithelial to mesenchymal transition (EMT) has been proposed to contribute to the metastatic spread of breast cancer cells. EMT-promoting transcription factors determine a continuum of different EMT states. In contrast, estrogen receptor α (ERα) helps to maintain the epithelial phenotype of breast cancer cells and its expression is crucial for effective endocrine therapies. Determining whether and how EMT-associated transcription factors such as ZEB1 modulate ERα signaling during early stages of EMT could promote the discovery of therapeutic approaches to suppress metastasis. Here we show that, shortly after induction of EMT and while cells are still epithelial, ZEB1 modulates ERα-mediated transcription induced by estrogen or cAMP signaling in breast cancer cells. Based on these findings and our ex vivo and xenograft results, we suggest that the functional interaction between ZEB1 and ERα may alter the tissue tropism of metastatic breast cancer cells towards bone.

Go to