Title

EZHIP Regulation in Cancer as a Therapeutic Strategy

Alternative Title

EZHIP 조절 기반 암 치료 전략

DGIST Authors

Jimin Moon ; Chan Chung ; Younghoon Kee

Advisor

정찬

Co-Advisor(s)

Younghoon Kee

Issued Date

2026

Awarded Date

2026-02-01

Type

Thesis

Description

EZHIP, Metformin, H3K27me3, KDM6, Osteosarcoma, Epigenetic Regulation, HTS|EZHIP, 메트포르민, H3K27me3, KDM6, 골육종, 후성유전학, 고속 스크리닝(HTS)

Table Of Contents

Ⅰ. Introduction 1
1. The Mechanism of EZHIP in Inhibiting PRC2 Methyltransferase Activity 1
2. The Contribution of EZHIP to the Aggressive Phenotype of Osteosarcoma 2
3. EZHIP-Associated Pathological Features of Pediatric Posterior Fossa Ependymoma (PFA-EPN) 3
4. Metformin-Mediated Modulation of EZHIP and H3K27me3 in PFA-EPN 4
5. Metformin-Dependent Regulation of EZHIP and the KDM6 Family 5
6. Metformin-Mediated Regulation of DNA Methylation 6
7. High-Throughput Screening of PTM-Dependent Modulators of EZHIP 7

ⅠⅠ. Results 8
1. The Effect of Metformin on EZHIP 8
1.1. Establishment of EZHIP Model System and Analysis of its Effects on Proliferation and Epigenetics 8
1.2. Metformin-Mediated Reduction of EZHIP, Restoration of H3K27me3, and Suppression of Cell Viability 13

2. EZHIP Suppression by Metformin Independent of Protein Degradation and DNA Methylation 18

3. Regulation of EZHIP by Metformin via Histone Methylation 22
3.1. Metformin-Induced Histone Modifications 22
3.2. Lack of Regulatory Contribution of H3K27ac to EZHIP Expression 24
3.3. Metformin-Mediated Modulation of KDM6 Leading to Elevated H3K27me3 and Downregulation of EZHIP 25

4. In vivo suppression of U2OS tumors by metformin 30

5. High-Throughput Screening–Based Identification of Kinase Activation as PTM Regulators of EZHIP 33

ⅠⅠⅠ. Discussion 40

ⅠV. Materials and Methods 43
1. Cell culture 43
2. Plasmid transformation 43
3. Stable Cell Line Generation 44
4. siRNA-mediated gene knockdown 45
5. shRNA-mediated gene knockdown 45
6. CCK-8 viability assay 46
7. Cell counting and viability analysis 46
8. RNA extraction 47
9. cDNA synthesis 47
10. Quantitative reverse transcription PCR 48
11. Whole cell lysate 50
12. Histone extraction 50
13. Western blotting 51
14. Immunofluorescence 52
15. Bisulfite sequencing 53
16. Chromatin Immunoprecipitation–quantitative PCR 54
17. Subcutaneous injection for xenograft mouse model 56
18. Tumor measurement 57
19. Intraperitoneal (IP) injection of metformin 57
20. High-throughput Screening 58

References 59

Summary (Korean) 62

URI

https://scholar.dgist.ac.kr/handle/20.500.11750/59656
http://dgist.dcollection.net/common/orgView/200000942420

DOI

10.22677/THESIS.200000942420

Degree

Master

Department

Department of New Biology

Publisher

DGIST

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