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DNA methylation analysis of human multiple myeloma.January 2006 (has links)
Cheung Kin Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 87-105). / Abstracts in English and Chinese. / Abstract (English version) --- p.i / Abstract (Chinese version) --- p.iii / Acknowledgments --- p.vi / Table of Contents --- p.v / List of Tables --- p.viii / List of Figures --- p.iv / List of Abbreviations --- p.xi / Chapter CHAPTER 1 --- GENERAL INTRODUCTION --- p.1 / Chapter CHAPTER 2 --- LITERATURE REVIEW --- p.3 / Chapter 2.1 --- Multiple myeloma --- p.3 / Chapter 2.2 --- Epidemiology of MM --- p.3 / Chapter 2.3 --- Risk factors --- p.4 / Chapter 2.4 --- Pathophysiology of MM --- p.5 / Chapter 2.5 --- Clinical presentations and diagnosis --- p.6 / Chapter 2.5.1 --- Diagnosis --- p.6 / Chapter 2.5.1.1 --- Laboratory testing of blood and urine --- p.6 / Chapter 2.5.1.2 --- Radiographic evaluations --- p.1 / Chapter 2.5.1.3 --- Bone marrow biopsy --- p.7 / Chapter 2.6 --- Staging and classification --- p.9 / Chapter 2.6.1 --- Staging --- p.9 / Chapter 2.6.2 --- Classification --- p.11 / Chapter 2.6.2.1 --- Monoclonal gammopathy of undetermined significance --- p.11 / Chapter 2.6.2.2 --- Asymptomatic MM --- p.12 / Chapter 2.6.2.3 --- Smouldering MM --- p.12 / Chapter 2.6.2.4 --- Indolent MM --- p.12 / Chapter 2.6.2.5 --- Symptomatic MM --- p.12 / Chapter 2.7 --- Treatment --- p.14 / Chapter 2.8 --- Epigenetics: DNA methylation --- p.15 / Chapter 2.9 --- Fundamental aspects of DNA methylation --- p.16 / Chapter 2.9.1 --- CpG islands --- p.16 / Chapter 2.9.2 --- Roles of DNA methylation --- p.16 / Chapter 2.9.3 --- Proposed mechanisms of transcriptional repression mediated by methylation --- p.18 / Chapter 2.10 --- Possible mechanisms to initiate aberrant DNA methylation --- p.21 / Chapter 2.11 --- DNA methylation in tumorigenesis --- p.22 / Chapter 2.11.1 --- Oncogenic point C → T mutation --- p.22 / Chapter 2.11.2 --- Global DNA hypomethylation --- p.23 / Chapter 2.11.3 --- Regional DNA hypermethylation --- p.23 / Chapter 2.12 --- Aberrant DNA methylation in MM --- p.25 / Chapter 2.12.1 --- Self-sufficiency in growth signals --- p.25 / Chapter 2.12.2 --- Evading apoptosis --- p.26 / Chapter 2.12.3 --- Insensitivity to antigrowth signals --- p.26 / Chapter 2.12.4 --- Tissue invasion and metastasis --- p.27 / Chapter 2.12.5 --- Infinite replicative potential --- p.28 / Chapter 2.12.6 --- Genome instability --- p.30 / Chapter 2.13 --- Methodologies of DNA methylation analysis --- p.32 / Chapter 2.13.1 --- Genome wide screening method: MS.AP-PCR --- p.32 / Chapter 2.13.2 --- Combined bisulfite restriction analysis --- p.34 / Chapter 2.13.3 --- Cloned bisulfite genomic sequencing --- p.36 / Chapter 2.13.4 --- Treatment with demethylating agent --- p.36 / Chapter CHAPTER 3 --- MATERIALS AND METHODS --- p.38 / Chapter 3.1 --- MM specimens --- p.38 / Chapter 3.1.1 --- MM samples --- p.38 / Chapter 3.1.2 --- MM cell lines --- p.38 / Chapter 3.2 --- Magnetic cell sorting of CD138-positive plasma cells --- p.39 / Chapter 3.3 --- Isolation of nuclear pellet from PB --- p.41 / Chapter 3.4 --- "DNA extraction from MM cell lines, MM plasma cells and PB" --- p.41 / Chapter 3.5 --- MS.AP-PCR --- p.42 / Chapter 3.5.1 --- Restriction enzyme digestion of genomic DNA --- p.42 / Chapter 3.5.2 --- Arbitrarily primed polymerase chain reaction --- p.42 / Chapter 3.5.3 --- Isolation of differentially methylated DNA fragments --- p.43 / Chapter 3.6 --- Cloning of differentially methylated DNA fragments --- p.46 / Chapter 3.6.1 --- TA cloning --- p.46 / Chapter 3.6.2 --- Heat shock transformation --- p.46 / Chapter 3.6.3 --- Screening of positive clones by PCR --- p.46 / Chapter 3.6.4 --- Alkaline lysis for plasmid DNA preparation --- p.47 / Chapter 3.7 --- MS.AP-PCR sequence analysis --- p.47 / Chapter 3.7.1 --- Nucleotide sequencing --- p.47 / Chapter 3.7.2 --- CpG islands analysis of differentially methylated sequences --- p.48 / Chapter 3.8 --- DNA methylation analysis --- p.48 / Chapter 3.8.1 --- Sodium bisulfite modification --- p.48 / Chapter 3.8.2 --- Combined bisulfite restriction analysis --- p.49 / Chapter 3.8.3 --- Cloned bisulfite genomic sequencing --- p.49 / Chapter 3.9 --- Gene expression analysis --- p.50 / Chapter 3.9.1 --- RNA extraction --- p.50 / Chapter 3.9.2 --- Reverse transcription PCR --- p.50 / Chapter 3.9.3 --- 5'-aza-2'-deoxycytidine treatment --- p.51 / Chapter CHAPTER 4 --- RESULTS --- p.53 / Chapter 4.1 --- Generation of DNA methylation patterns by MS.AP-PCR --- p.53 / Chapter 4.1.1. --- Global methylation content in MM samples and normal PB lymphocytes --- p.56 / Chapter 4.1.2. --- Differential methylation in MM --- p.56 / Chapter 4.2 --- UCSC BLAT analysis of differentially methylated DNA fragments --- p.60 / Chapter 4.3 --- Identification of two candidate genes with downregulated expression --- p.60 / Chapter 4.4 --- Zinc fingers and homeoboxes 2 (ZHX2) --- p.62 / Chapter 4.4.1 --- ZHX2 CpG islands BLAT search analysis --- p.62 / Chapter 4.4.2 --- Hypermethylation of ZHX2 in MM cell lines --- p.63 / Chapter 4.4.3 --- Downregulated expression of ZHX2 in methylated MM cell lines --- p.66 / Chapter 4.4.4 --- Restoration of ZHX2 expression by 5-Aza-dC treatment --- p.67 / Chapter 4.4.5 --- Unmethylation of ZHX2 in primary MM tumors --- p.68 / Chapter 4.5 --- Ring finger protein 180 (RNF180) --- p.69 / Chapter 4.5.1 --- RNF180 CpG islands BLAT search analysis --- p.69 / Chapter 4.5.2 --- Hypermethylation of RNF180 in MM cell lines --- p.70 / Chapter 4.5.3 --- Downregulated expression of RNF180 in methylated MM cell lines --- p.73 / Chapter 4.5.4 --- Restoration of RNF180 expression by 5-Aza-dC treatment --- p.74 / Chapter 4.5.5 --- Methylation of RNF180 in primary MM tumors --- p.75 / Chapter CHAPTER 5 --- DISCUSSION --- p.76 / Chapter 5.1 --- Importance of methylation in MM --- p.76 / Chapter 5.2 --- Genome-wide screening approach by MS.AP-PCR --- p.76 / Chapter 5.3 --- Sample selection in MS.AP-PCR --- p.78 / Chapter 5.4 --- Methylation patterns in MM --- p.79 / Chapter 5.5 --- Candidate genes selection strategies --- p.81 / Chapter 5.6 --- Zinc fingers and homeoboxes 2 --- p.81 / Chapter 5.7 --- Ring finger protein 180 --- p.83 / Chapter 5.8 --- Limitations --- p.84 / Chapter CHAPTER 6 --- CONCLUSION --- p.86 / REFERENCES --- p.87
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A gene hypermethylation profile of non-astrocytic gliomas. / CUHK electronic theses & dissertations collectionJanuary 2002 (has links)
Dong Shumin. / "February 2002." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (p. 187-220). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.
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