Regulation of cancer progression by hepatitis B virus X protein (HBx) and HBxAP/RSF1

I. Regulation of catalase on hepatitis B virus X protein (HBx) in HBV-related advanced hepatocellular carcinomas 1 A. INTRODUCTION 2 B. MATERIALS AND METHODS. 4 1. Cell cultures and plasmids 4 2. RNA extraction, reverse transcription-polymerase chain reaction 4 3. Western blotting . 5 4. Luciferase assay 5 5. Clonogenic cell proliferation assays 5 6. Patients characteristics 6 7. Statistics 6 C. RESULTS 7 1. The protein levels of HBx are decreased by catalase and MnSOD 7 2. Catalase downregulates HBx protein levels at the translational level 7 3. Cysteine residues of HBx are essential to maintain protein stability . 9 4. Cysteine null mutant of HBx is insufficient for transactivation activities 12 5. Cysteine null mutant is not affected by catalase or N-acetyl cysteine 12 6. Cysteine residues are important for HBx-mediated clonogenic cell proliferation 14 7. Catalase expression level is lower in tumor tissues than in non-tumor tissues in HBV-related HCC. 17 8. Catalase expression level is significantly lower in HBV-related advanced HCC 17 9. HBx protein levels are negatively correlated with catalase expression in HBV-related advanced HCC. 20 10. HBx protein levels have no correlation with MnSOD expression in HBV-related advanced HCC 23 11. Prognostic significance of catalase expression in HBV-related advanced HCC 23 D. DISCUSSION 30 II. HBxAP/RSF1 as a chromatin remodeller and HBx-associated protein 32 A. INTRODUCTION 33 B. MATERIALS AND METHODS . 36 1. Cell cultures and antibody. 36 2. Plasmid. 36 3. Chromosome spreading and Giemsa staining. 36 4. FRET (Fluorescence Resonance Energy Transfer) assay. 37 5. Modified histone peptide array. 37 6. Purification of RSF1. 37 7. Protein microchip array. . 37 C. RESULTS 39 1. RSF1 is overexpressed in HBV-related advanced HCC. 39 2. RSF1 depletion induces premature separation of sister chromatids. 39 3. The centromeric expression level of H3K9me3 is downregulated at metaphase in RSF1 knock-down cells 43 4. RSF1 depletion enhances the expression levels of H3K9me3 and H3K27me3 in mitosis 46 5. RSF1 dissociates from histone peptides with phosphorylated H3S28 modification 46 6. RSF1 directly interacts with diverse targets. 48 D. DISCUSSION 51 III. CONCLUSION 53 IV. REFFERENCE 54 국문요약 59

Author dc:creator

• 조, 미영

Contributors dc:contributor

• 조, 혜성
• 대학원 의생명과학과
• 110686

• Hepatitis B virus X (HBx)
• catalase
• cysteine
• liver cancer
• HBxAP/RSF1

Language dc:language

en

Identifier

http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000021592
000000021592

OAI identifier oai:identifier

oai:repository.ajou.ac.kr:201003/13015