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Updated: May 7, 2026
Systems Biology of Metabolic Regulation by Estrogen Receptor Signaling in Breast Cancer
Published on: March 17, 2016
Distinct pathways regulated by RET and estrogen receptor in luminal breast cancer demonstrate the biological basis
Philip M Spanheimer1, Anthony R Cyr, Matthew P Gillum
1From the Department of Surgery, University of Iowa, Iowa City.
Objective:
We investigated directed therapy based on TFAP2C-regulated pathways to inform new therapeutic approaches for treatment of luminal breast cancer.
Background:
TFAP2C regulates the expression of genes characterizing the luminal phenotype including ESR1 and RET, but pathway cross talk and potential for distinct elements have not been characterized.
Methods:
Activation of extracellular signal-regulated kinases (ERK) and AKT was assessed using phosphorylation-specific Western blot. Cell proliferation was measured with MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] after siRNA (small interfering RNA) gene knockdown or drug treatment. Cell cycle, Ki-67, and cleaved caspase 3 were measured by fluorescence-activated cell sorting. Tumorigenesis was assessed in mice xenografts.
Results:
Knockdown of TFAP2C or RET inhibited GDNF (glial cell line-derived neurotrophic factor)-mediated activation of ERK and AKT in MCF-7 cells. Similarly, sunitinib, a small-molecule inhibitor of RET, blocked GDNF-mediated activation of ERK and AKT. Inhibition of RET either by gene knockdown or by treatment with sunitinib or vandetanib reduced RET-dependent growth of luminal breast cancer cells. Interestingly, knockdown of TFAP2C, which controls both ER (estrogen receptor) and RET, demonstrated a greater effect on cell growth than either RET or ER alone. Parallel experiments using treatment with tamoxifen and sunitinib confirmed the increased effectiveness of dual inhibition of the ER and RET pathways in regulating cell growth. Whereas targeting the ER pathway altered cell proliferation, as measured by Ki-67 and S-phase, anti-RET primarily increased apoptosis, as demonstrated by cleaved caspase 3 and increased TUNEL (terminal deoxyneucleotidyl transferase dUTP nick end labeling) expression in xenografts.
Conclusions:
ER and RET primarily function through distinct pathways regulating proliferation and cell survival, respectively. The findings inform a therapeutic approach based on combination therapy with antiestrogen and anti-RET in luminal breast cancer.
Insights
Targeting both estrogen receptor (ER) and RET pathways with combination therapy shows promise for luminal breast cancer treatment. Dual inhibition of ER and RET pathways significantly impacts cell growth and survival.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Therapeutics
Background:
- TFAP2C (Transcription Factor AP-2 gamma) is crucial for luminal breast cancer phenotype, regulating ESR1 and RET.
- The crosstalk between TFAP2C-regulated pathways and their distinct roles in cancer progression require further characterization.
Purpose of the Study:
- To investigate TFAP2C-regulated pathways for novel therapeutic strategies in luminal breast cancer.
- To elucidate the distinct roles of ER and RET pathways in regulating cancer cell proliferation and survival.
Main Methods:
- Utilized siRNA gene knockdown and small-molecule inhibitors (sunitinib, vandetanib) to target TFAP2C, ER, and RET.
- Assessed extracellular signal-regulated kinases (ERK) and AKT activation via Western blot.
- Measured cell proliferation, cell cycle, apoptosis (cleaved caspase 3, TUNEL), and tumorigenesis in mouse xenografts.
Main Results:
- TFAP2C or RET inhibition, and sunitinib treatment, blocked GDNF-mediated ERK and AKT activation.
- Targeting RET reduced RET-dependent luminal breast cancer cell growth.
- Dual inhibition of ER and RET pathways demonstrated superior efficacy in controlling cell growth compared to targeting either pathway alone.
- ER pathway inhibition primarily affected proliferation, while RET inhibition induced apoptosis.
Conclusions:
- Estrogen receptor (ER) and RET pathways operate distinctly, regulating proliferation and cell survival, respectively.
- Combination therapy targeting both ER and RET pathways offers a promising therapeutic approach for luminal breast cancer.
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