Novel Functions of EZH2 in Triple Negative Breast Cancer: Translation in to New Biomarker and Treatment Strategies
Abstract
In our third year of funding we have developed an antibody that recognizes the cytoplasmic phosphorylated EZH2 protein (pEZH2(T367)), and validated the specificity in human tissue samples by immunohistochemistry. Using this novel antibody, we found that pEZH2 (T367) expression in the cytoplasm increases with breast cancer progression, and that it is significantly inversely associated with H3K27me3 in breast cancer tissue samples. We have completed our binding assays using EZH2 and p38 proteins by quantitative bio-layer interferometry (BLI) and conclusively demonstrated a strong binding affinity of EZH2 for p38 (binding affinity KD of 5.54 nM). We successfully optimized a proximity ligation assay (PLA) to investigate the binding of these proteins in situ. This assay confirmed that EZH2 binds to p38 in breast cancer cell lines and that the binding occurs predominantly in the nucleus of breast cancer cells, which was previously unknown. We have developed various EZH2 mutants that will be helpful in identifying the required EZH2-p38 binding sites.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 2018
- Accession Number
- AD1063037
Entities
People
- Zaneta Nikolovska-coleska
Organizations
- University of Michigan