Molecular study of the terminal differentiation of WEHI-3B JCS myeloid leukemia cell induced by biochanin A.
1998
Nachschlagewerk
Zugriff:
by Yip Mei Chu Pandora.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1998.
Includes bibliographical references (leaves 207-233).
also in Chinese.
STATEMENT --- p.i
ACKNOWLEDGEMENTS --- p.ii
p.iii
(CHINESE VERSION) --- p.v
TABLE OF CONTENTS --- p.vii
ABBREVIATIONS --- p.xiii
LIST OF FIGURES AND TABLES --- p.xvii
Chapter CHAPTER ONE ... --- GENERAL INTRODUCTION
Chapter 1.1 --- the blood cells formation - hematopoiesis --- p.1
Chapter 1.1.1 --- Hierarchy of hematopoiesis --- p.2
Chapter 1.1.2 --- Malfunction in the process of hematopoiesis - hematologic neoplasia - Leukemia --- p.6
Chapter 1.1.2.1 --- Classification of leukemia --- p.7
Chapter 1.1.2.2 --- Differentiation therapy ´ؤ a new hope in the treatment of leukemia --- p.9
Chapter 1.2 --- Understanding the pathogenesis of leukemia --- p.12
Chapter 1.2.1 --- General regulation of hematopoiesis --- p.12
Chapter 1.2.2 --- Regulation of the differentiation of myeloid lineage --- p.15
Chapter 1.2.2.1 --- Regulation of myeloid cell differentiation by hematopoietic regulatory protein --- p.16
Chapter 1.2.2.2 --- Signal transduction pathways in myeloid cell differentiation --- p.20
Chapter 1.2.2.3 --- Gene regulation of myeloid cell differentiation --- p.22
Chapter 1.2.2.3.1 --- Transcription factors --- p.23
Chapter 1.2.2.3.2 --- Myeloid specific genes --- p.31
Chapter 1.2.2.3.3 --- Protooncogenes and tumor suppressor genes --- p.37
Chapter 1.2.2.3.4 --- Homeobox genes --- p.42
Chapter 1.2.2.3.5 --- Cell cycle control in myeloid growth and differentiation --- p.47
Chapter 1.3 --- Induction of differentiation in myeloid leukemia cell --- p.48
Chapter 1.3.1 --- Induced myeloid leukemia cell differentiation --- p.48
Chapter 1.3.2 --- Inducers of myeloid cell differentiation --- p.52
Chapter 1.3.3 --- Chemical inducers ´ؤ Flavonoids --- p.57
Chapter 1.3.4 --- Murine myeloid leukemia cell ´ؤ WEHI-3B JCS --- p.60
Chapter 1.4 --- Aim of study --- p.53
Chapter CHAPTER TWO ... --- ISOLATION OF GENES THAT ARE DIFFERENTIALLY EXPRESSED DURING BIOCHANIN A INDUCED WEHI-3B (JCS) MYELOID LEUKEMIA CELL DIFFERENTIATION
Chapter 2.1 --- Introduction --- p.65
Chapter 2.1.1 --- Strategy for searching differentially expressed genes - RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP- PCR) --- p.65
Chapter 2.1.2 --- Reamplification of PCR products by Touchdown PCR --- p.67
Chapter 2.1.3 --- Methods for eliminating false positives : Dot blot hybridization screening --- p.68
Chapter 2.2 --- Materials --- p.70
Chapter 2.2.1 --- "Cell line, Bacterial strain and Vector" --- p.70
Chapter 2.2.2 --- Chemicals --- p.70
Chapter 2.2.3 --- Reagents and nucleic acids --- p.71
Chapter 2.2.4 --- Kits --- p.72
Chapter 2.2.5 --- Solutions --- p.72
Chapter 2.2.6 --- Equipments --- p.73
Chapter 2.3 --- Methods --- p.74
Chapter 2.3.1 --- Induction of murine myeloid leukemia cell line -WEHI-3B (JCS) cells by biochanin-A --- p.74
Chapter 2.3.2 --- Isolation of total RNA by guanidium thiocyanate cesium chloride ultracentrifugation --- p.74
Chapter 2.3.3 --- RNA fingerprinting by arbitrarily primed PCR --- p.75
Chapter 2.3.3.1 --- Synthesis of first strand cDNA --- p.75
Chapter 2.3.3.2 --- Normalization of RNA samples --- p.75
Chapter 2.3.3.3 --- RAP-PCR --- p.76
Chapter 2.3.3.4 --- Reamplification of differentially amplified fragment --- p.77
Chapter 2.3.4 --- First round dot blot hybridization screening --- p.78
Chapter 2.3.4.1 --- Dot blot --- p.78
Chapter 2.3.4.2 --- Preparation of cDNA probe --- p.79
Chapter 2.3.4.3 --- 32P-labelling of cDNA probe --- p.79
Chapter 2.3.4.4 --- Removal of unincorporated probe by NICK´ёØ column --- p.80
Chapter 2.3.4.5 --- Estimation of 32P labelling efficiency by scintillation counting --- p.80
Chapter 2.3.4.6 --- Prehybridization and hybridization --- p.81
Chapter 2.3.4.7 --- Quantitation of hybridization signal by scanning densitometry --- p.81
Chapter 2.3.5 --- Second round dot blot hybridization screening --- p.81
Chapter 2.3.5.1 --- Subcloning of differentially amplified fragments --- p.82
Chapter 2.3.5.1.1 --- Preparation of vector DNA --- p.82
Chapter 2.3.5.1.2 --- Synthesis of blunt end PCR product --- p.84
Chapter 2.3.5.1.3 --- Blunt end ligation --- p.34
Chapter 2.3.5.1.4 --- Transformation --- p.85
Chapter 2.3.5.1.5 --- Selection and confirmation by polymerase chain reaction --- p.85
Chapter 2.3.5.2 --- Dot blot hybridization screening --- p.85
Chapter 2.4 --- Results --- p.87
Chapter 2.4.1 --- Spectrophotometric analysis of total RNA --- p.87
Chapter 2.4.2 --- Normalization of RNA samples --- p.88
Chapter 2.4.3 --- RNA fingerprinting by arbitrarily primed PCR --- p.39
Chapter 2.4.4 --- Reamplification of isolated RAP-PCR products --- p.91
Chapter 2.4.5 --- First round of dot blot hybridization screening --- p.92
Chapter 2.4.6 --- Subcloning of differentially amplified fragments --- p.100
Chapter 2.4.7 --- Second round of dot blot hybridization screening --- p.102
Chapter 2.4.8 --- Comparison of the first and second round of dot blot hybridization screening --- p.106
Chapter 2.5 --- Discussion --- p.108
Chapter 2.5.1 --- RNA fingerprinting by arbitrarily primed PCR --- p.108
Chapter 2.5.2 --- Limitation of RAP-PCR --- p.110
Chapter 2.5.3 --- Two rounds of dot blot hybridization screening --- p.111
Chapter CHAPTER THREE... --- CHARACTERIZATION OF THE ISOLATED GENE FRAGMENTS
Chapter 3.1 --- Introduction --- p.113
Chapter 3.1.1 --- Automated DNA sequencing and analysis --- p.113
Chapter 3.1.2 --- GenBank and the BLAST homology search --- p.115
Chapter 3.2 --- Materials --- p.118
Chapter 3.2.1 --- Selected recombinant plasmids --- p.118
Chapter 3.2.2 --- Chemicals --- p.118
Chapter 3.2.3 --- Reagents --- p.118
Chapter 3.2.4 --- Kits --- p.119
Chapter 3.2.5 --- Solutions --- p.119
Chapter 3.2.6 --- Equipment --- p.119
Chapter 3.3 --- Methods --- p.120
Chapter 3.3.1 --- Preparation of selected recombinant plasmid DNA --- p.120
Chapter 3.3.2 --- Restriction digestion of recombinant plasmid DNA --- p.120
Chapter 3.3.3 --- Automated DNA sequencing --- p.120
Chapter 3.3.3.1 --- Primer annealing to template --- p.120
Chapter 3.3.3.2 --- Sequencing reactions --- p.121
Chapter 3.3.3.3 --- Polyacrylamide gel electrophoresis --- p.121
Chapter 3.3.3.4 --- Data analysis by ALF manager and DNAsis --- p.122
Chapter 3.3.4 --- Sequence homology search with databases --- p.122
Chapter 3.4 --- Results --- p.123
Chapter 3.4.1 --- Spectrophotometric analysis of selected recombinant plasmid DNAs subcloned with differentially amplified fragments --- p.123
Chapter 3.4.2 --- Restriction digestion of selected recombinant plasmid DNA --- p.124
Chapter 3.4.3 --- Sequences of the subcloned differentially amplified fragments --- p.126
Chapter 3.4.4 --- Sequence analysis of the subcloned differentially amplified fragments --- p.144
Chapter 3.5 --- Discussion --- p.157
Chapter 3.5.1 --- Sequence analysis of the isolated gene fragment --- p.157
Chapter CHAPTER FOUR … --- "EXPRESSION PROFILE OF ISOLATED GENES FRAGMENTS IN MYELOID LEUKEMIA CELL, MOUSE EMBRYO, AND TISSUES"
Chapter 4.1 --- Introduction --- p.162
Chapter 4.1.1 --- Quantitation of mRNA by Reverse transcription-polymerase chain reaction --- p.162
Chapter 4.1.2 --- Internal primer design by OLIGO´ёØ ver 34 --- p.167
Chapter 4.2 --- Materials --- p.168
Chapter 4.2.1 --- Mice --- p.168
Chapter 4.2.2 --- Cell lysate --- p.168
Chapter 4.2.3 --- Total RNAs --- p.168
Chapter 4.3 --- Methods --- p.169
Chapter 4.3.1 --- Internal primer design by OLIGO´ёØ ver 34 --- p.169
Chapter 4.3.2 --- "Isolation of total RNA from biochanin A induced JCS cells, mouse embryos and tissue" --- p.169
Chapter 4.3.2.1 --- Preparation of cell lysate from mouse embryo and postnatal mouse brain --- p.169
Chapter 4.3.2.2 --- Isolation of RNA by guanidium thiocyanate cesium chloride method --- p.170
Chapter 4.3.3 --- Preparation of saggital section of mouse embryo --- p.170
Chapter 4.3.4 --- Confirmation of differential expression of isolated genes fragments during biochanin A and midazolam induced WEHI 3B (JCS) differentiation and the expression profile in mouse tissues and during mouse embryo development by reverse transcription-polymerase chain reaction --- p.171
Chapter 4.4 --- Results --- p.173
Chapter 4.4.1 --- Internal primer design of the sequenced fragments --- p.173
Chapter 4.4.2 --- Spectrophotometric analysis of total RNA --- p.175
Chapter 4.4.3 --- Saggital section of mouse embryo --- p.176
Chapter 4.4.4 --- Normalization of RNA samples --- p.180
Chapter 4.4.5 --- Analysis of mRNA expression of differentially amplified fragmentsin biochanin A or midazolam induced JCS cells and mouse embryos by RT- PCR --- p.182
Chapter 4.4.5.1 --- "Genes downregulated at 1 hour, 5 hours and 48 hours after biochanin A induction of JCS cells" --- p.183
Chapter 4.4.5.2 --- Genes up-regulated at 48 hours after biochanin A induction --- p.183
Chapter 4.4.5.3 --- Genes constitutively expressed during the course of biochanin A treatment --- p.184
Chapter 4.4.5.4 --- Genes showing undetectable level of expression in biochanin A induced JCS cells --- p.184
Chapter 4.4.6 --- Tissue expression of the biochanin A induced-differentially expressed fragments by RT-PCR --- p.188
Chapter 4.5 --- Discussion --- p.191
Chapter 4.5.1 --- Expression profiles of isolated differentially amplified fragments --- p.191
Chapter 4.5.2 --- Comparison of the expression profiles of the isolated gene fragments analyzed by dot blot hybridization screening and RT-PCR --- p.197
Chapter CHAPTER FIVE ... --- GENERAL DISCUSSION --- p.200
REFERENCES --- p.207
APPENDIX --- p.234
| Titel: |
Molecular study of the terminal differentiation of WEHI-3B JCS myeloid leukemia cell induced by biochanin A.
|
|---|---|
| Autor/in / Beteiligte Person: | Yip, Mei Chu Pandora. ; Chinese University of Hong Kong Graduate School. Division of Biology. |
| Link: | |
| Veröffentlichung: | 1998 |
| Medientyp: | Nachschlagewerk |
| Schlagwort: |
|
| Sonstiges: |
|