Jiwon Hwang. (2025). EZH Inhibitory Protein (EZHIP) Regulation by Targeting Genetic and Epigenetic Alterations. doi: 10.22677/THESIS.200000844106
Type
Thesis
Description
EZHIP, Metformin, H3K27me3, KDM6, Cancer Epigenetics
Table Of Contents
Abstract List of Contents Abbreviations List of Tables List of Figures
Ⅰ. Introduction 1. EZHIP overexpression inhibits PRC2 in various cancers 2. PFA Ependymoma is a fatal pediatric brain tumor 3. Metformin treatment reverses EZHIP expression and restores H3K27me3 levels 4. KDM6 is the potential target of metformin treatment
ⅠⅠ. Results 1. Effect of metformin on EZHIP-expressing cells 1.1. EZHIP levels are increased in U2OS 1.2. Metformin is cytotoxic to EZHIP-expressing cell lines 1.3. Metformin induces EZHIP reduction and H3K27me3 upregulation in EZHIP-expressing cells 1.4. Metformin alters EZHIP expression at the transcriptional level
2. Regulation of EZHIP by DNA Methylation 2.1. DNA demethylation induces EZHIP expression 2.2. Endogenous EZHIP expression is determined by DNA methylation 2.3. Metformin leads to subtle methylation in the EZHIP gene body
3. Regulation of EZHIP via KDM6 Family Reduction by Metformin 3.1. Metformin interacts with the histone demethylase KDM6 family 3.2. Metformin downregulates KDM6 family 3.3. KDM6A and KDM6B are potential targets for EZHIP reduction
ⅠⅠⅠ. Discussion
ⅠV. Conclusion and Future Perspective
V. Method and Materials 1. Cell Culture 2. Cell Viability Assay 3. Histone Extraction 4. Western Blotting 5. Quantitative RT-PCR (RT-qPCR) 6. Bisulfite sequencing 7. Chromatin Immunoprecipitation RT-qPCR (ChIP-qPCR) 8. Transformation and Plasmid Preparation 9. Stable Cell Line Generation 10. siRNA Transfection 11. Quantification and Statistical Analysis
