Elevated GRHL2 Imparts Plasticity in ER-Positive Breast Cancer Cells
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Generation of Tetracycline-Inducible GRHL2-GFP Expression in MCF7 Cell Line
2.3. Drug Treatment
2.4. RNA Isolation and RT-qPCR
2.5. RNA Sequencing
2.6. Protein Preparation and Western Blotting
2.7. Chromatin Immunoprecipitation (ChIP) and Sequencing
2.8. Transient Transfection
2.9. Migration Assay
2.10. Flow Cytometry
2.11. Immunocytochemistry and Immunohistrochemistry
2.12. Soft Agar Assay
2.13. Mammosphere Assay
2.14. Tumor Xenograft
2.15. Statistical Analysis
3. Results
3.1. A Tetracycline-Inducible Model of High GRHL2 Expression in Breast Cancer Cells Expresses GRHL2 Protein and mRNA in a Dox Dose-Dependent Manner
3.2. High GRHL2 Expression Increases Epithelial Cell Phenotypes
3.3. GRHL2 Overexpression Alters Its Endogenous Transcriptional Activity and Gene Expression
3.4. GRHL2 Overexpression Regulates Growth and Developmental Pathways
3.5. GRHL2 Overexpression Inhibits Proliferation in Vivo
3.6. GRHL2 Overexpression Enriches Stem Cell-like Characteristics
3.7. GRHL2 Overexpression Leads to a Complex Epithelial–Mesenchymal Hybrid Phenotype
3.8. GRHL2 Overexpression Alters GRHL2 Genome Binding in a Dynamic Manner
3.9. GRHL2 Overexpression Changes Motifs Found near GRHL2 Binding Sites
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Zheng, C.; Allen, K.O.; Liu, T.; Solodin, N.M.; Meyer, M.B.; Salem, K.; Tsourkas, P.K.; McIlwain, S.J.; Vera, J.M.; Cromwell, E.R.; et al. Elevated GRHL2 Imparts Plasticity in ER-Positive Breast Cancer Cells. Cancers 2024, 16, 2906. https://doi.org/10.3390/cancers16162906
Zheng C, Allen KO, Liu T, Solodin NM, Meyer MB, Salem K, Tsourkas PK, McIlwain SJ, Vera JM, Cromwell ER, et al. Elevated GRHL2 Imparts Plasticity in ER-Positive Breast Cancer Cells. Cancers. 2024; 16(16):2906. https://doi.org/10.3390/cancers16162906
Chicago/Turabian StyleZheng, Christy, Kaelyn O. Allen, Tianrui Liu, Natalia M. Solodin, Mark B. Meyer, Kelley Salem, Phillipos K. Tsourkas, Sean J. McIlwain, Jessica M. Vera, Erika R. Cromwell, and et al. 2024. "Elevated GRHL2 Imparts Plasticity in ER-Positive Breast Cancer Cells" Cancers 16, no. 16: 2906. https://doi.org/10.3390/cancers16162906
APA StyleZheng, C., Allen, K. O., Liu, T., Solodin, N. M., Meyer, M. B., Salem, K., Tsourkas, P. K., McIlwain, S. J., Vera, J. M., Cromwell, E. R., Ozers, M. S., Fowler, A. M., & Alarid, E. T. (2024). Elevated GRHL2 Imparts Plasticity in ER-Positive Breast Cancer Cells. Cancers, 16(16), 2906. https://doi.org/10.3390/cancers16162906