Epigenetic regulation of ID4 in breast cancer: tumor suppressor or oncogene?
Related Experiment Video
Updated: Feb 6, 2026
Orthotopic Transplantation of Breast Tumors as Preclinical Models for Breast Cancer
Published on: May 18, 2020
Daniela Nasif1, Emanuel Campoy2, Sergio Laurito3
1IHEM, National University of Cuyo, CONICET, Mendoza, Argentina.
Background:
Inhibitor of differentiation protein 4 (ID4) is a dominant negative regulator of the basic helix-loop-helix (bHLH) family of transcription factors. During tumorigenesis, ID4 may act as a tumor suppressor or as an oncogene in different tumor types. However, the role of ID4 in breast cancer is not clear where both an oncogenic and a tumor suppressor function have been attributed. Here, we hypothesize that ID4 behaves as both, but its role in breast differs according to the estrogen receptor (ER) status of the tumor.
Methods:
ID4 expression was retrieved from TCGA database using UCSC Xena. Association between overall survival (OS) and ID4 was assessed using Kaplan-Meier plotter. Correlation between methylation and expression was analyzed using the MEXPRESS tool. In vitro experiments involved ectopic expression of ID4 in MCF-7, T47D, and MDA-MB231 breast cancer cell lines. Migration and colony formation capacity were assessed after transfection treatments. Gene expression was analyzed by ddPCR and methylation by MSP, MS-MLPA, or ddMSP.
Results:
Data mining analysis revealed that ID4 expression is significantly lower in ER+ tumors with respect to ER- tumors or normal tissue. We also demonstrate that ID4 is significantly methylated in ER+ tumors. Kaplan-Meier analysis indicated that low ID4 expression levels were associated with poor overall survival in patients with ER+ tumors. In silico expression analysis indicated that ID4 was associated with the expression of key genes of the ER pathway only in ER+ tumors. In vitro experiments revealed that ID4 overexpression in ER+ cell lines resulted in decreased migration capacity and reduced number of colonies. ID4 overexpression induced a reduction in ER levels in ER+ cell lines, while estrogen deprivation with fulvestrant did not induce changes neither in ID4 methylation nor in ID4 expression.
Conclusions:
We propose that ID4 is frequently silenced by promoter methylation in ER+ breast cancers and functions as a tumor suppressor gene in these tumors, probably due to its interaction with key genes of the ER pathway. Our present study contributes to the knowledge of the role of ID4 in breast cancer.
Insights
Inhibitor of differentiation protein 4 (ID4) acts as a tumor suppressor in estrogen receptor-positive breast cancer, often silenced by methylation. Low ID4 expression correlates with poor survival in ER+ patients.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Inhibitor of differentiation protein 4 (ID4) is a transcription factor regulator with varied roles in tumorigenesis.
- Its function in breast cancer remains unclear, with evidence suggesting both oncogenic and tumor-suppressive activities.
- This study investigates ID4's dual role, hypothesizing it differs based on tumor estrogen receptor (ER) status.
Purpose of the Study:
- To elucidate the specific role of ID4 in breast cancer subtypes.
- To determine if ID4's function correlates with estrogen receptor (ER) positivity or negativity.
- To investigate the mechanisms of ID4 regulation, including methylation, in ER+ breast cancer.
Main Methods:
- Utilized TCGA database for ID4 expression analysis and correlated it with overall survival (OS) using Kaplan-Meier plotter.
- Analyzed ID4 methylation and expression correlation using MEXPRESS.
- Conducted in vitro experiments involving ID4 overexpression in ER+ (MCF-7, T47D) and ER- (MDA-MB231) breast cancer cell lines, assessing migration and colony formation.
- Quantified gene expression and methylation using ddPCR and MSP/MS-MLPA/ddMSP.
Main Results:
- ID4 expression was significantly lower in ER+ tumors compared to ER- tumors and normal tissue.
- ID4 promoter methylation was significantly higher in ER+ tumors, correlating with lower ID4 expression.
- Low ID4 expression was linked to poorer OS in ER+ breast cancer patients.
- ID4 overexpression in ER+ cell lines reduced migration and colony formation, and decreased ER levels.
- Estrogen deprivation did not affect ID4 methylation or expression.
Conclusions:
- ID4 functions as a tumor suppressor in ER+ breast cancer, frequently silenced by promoter methylation.
- ID4's suppressive role is likely mediated through interaction with ER pathway genes.
- This study clarifies ID4's context-dependent role in breast cancer, particularly in ER+ subtypes.
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