Molecular study of differentially expressed genes in prostaglandin E₂ induced WEHI-3B JCS-14 and JCS cell differentiation.
2003
Nachschlagewerk
Zugriff:
Chan Sin-Man.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2003.
Includes bibliographical references (leaves 154-169).
in English and Chinese.
Acknowledgements --- p.i
p.iv
(Chinese Version) --- p.vi
Contents --- p.viii
Abbreviations --- p.xiii
List of Figures and Tables --- p.xvi
Chapter Chapter One --- General Introduction
Chapter 1.1 --- Hematopoiesis --- p.1
Chapter 1.1.1 --- Ontogeny of hematopoiesis --- p.1
Chapter 1.1.2 --- Hiercharay of hematopoiesis --- p.2
Chapter 1.2 --- Regulation of hematopoiesis --- p.5
Chapter 1.2.1 --- Bone marrow stromal cell --- p.5
Chapter 1.2.2 --- Hematopoietic growth factor --- p.6
Chapter 1.2.3 --- Hematopoietic growth factor receptors and signal transduction --- p.10
Chapter 1.2.4 --- Transcriptional regulation of myeloid cell development --- p.11
Chapter 1.3 --- Deregulated hematopoiesis - Leukemia --- p.20
Chapter 1.3.1 --- Classification of leukemia --- p.20
Chapter 1.3.2 --- Molecular basis of leukemia --- p.20
Chapter 1.4 --- Prostaglandin E2 induced WEHI-3B JCS and JCS-14 cell differentiation --- p.22
Chapter 1.4.1 --- Induced leukemia cell differentiation --- p.22
Chapter 1.4.2 --- Inducer of cell differentiation - Prostaglandin E2 --- p.22
Chapter 1.4.3 --- WEHI-3B JCS and subline JCS-14 cells --- p.24
Chapter 1.5 --- The aims of study --- p.26
Chapter Chapter Two --- Identification of differentially expressed genes during PGE2-induced WEHI-3B JCS-14 cell differentiation
Chapter 2.1 --- Introduction --- p.27
Chapter 2.1.1 --- Strategy for studying PGE2-induced JCS-14 cell differentiation --- p.28
Chapter 2.1.2 --- Method for studying differential gene expression: Microarry Technology --- p.29
Chapter 2.2 --- Materials --- p.32
Chapter 2.2.1 --- Cell line --- p.32
Chapter 2.2.2 --- AtlasT M Mouse cDNA Expression Array --- p.32
Chapter 2.2.3 --- Chemicals --- p.32
Chapter 2.2.4 --- Solutions and buffers --- p.33
Chapter 2.2.5 --- Reagents --- p.34
Chapter 2.3 --- Methods --- p.35
Chapter 2.3.1 --- Morphological study of PGE2-induced JCS-14 cell differentiation --- p.35
Chapter 2.3.2 --- Preparation of total RNA from PGE2-induced JCS-14 cells --- p.35
Chapter 2.3.2.1 --- Preparation of cell lysates --- p.35
Chapter 2.3.2.2 --- Isolation of total RNA --- p.35
Chapter 2.3.3 --- Preparation of cDNA probes --- p.36
Chapter 2.3.3.1 --- Probe synthesis from total RNA --- p.36
Chapter 2.3.3.2 --- Purification of the labeled cDNA probes --- p.37
Chapter 2.3.4 --- Hybridization cDNA probes to the Atlas Array and stringency wash --- p.37
Chapter 2.4 --- Results --- p.39
Chapter 2.4.1 --- Morphological changes in PGE2-treated JCS-14 cells --- p.39
Chapter 2.4.2 --- Analysis of total RNA from PGE2-induced JCS-14 cells --- p.43
Chapter 2.4.3 --- Hybridization of cDNA probes to AtlasT M cDNA Expression Array --- p.45
Chapter 2.5 --- Discussion --- p.73
Chapter 2.5.1 --- Morphological study of JCS-14 cell differentiation --- p.73
Chapter 2.5.2 --- Differentiation commitment of JCS-14 cell under PGE2 induction --- p.73
Chapter 2.5.3 --- Gene expression profile by microarray --- p.74
Chapter 2.5.4 --- Gene expression profile of 5 hours PGE2-induced JCS-14 cells --- p.74
Chapter 2.5.5 --- Further analysis of regulatory genes in PGE2-induced JCS-14 cell differentiation --- p.77
Chapter Chapter Three --- Expression profile of identified genes in WEHI-3B JCS-14 and JCS cell differentiation
Chapter 3.1 --- Introduction --- p.79
Chapter 3.1.1 --- Quantitation of mRNA by Real time RT-PCR --- p.80
Chapter 3.1.2 --- Relative quantitation of gene expression --- p.83
Chapter 3.2 --- Materials --- p.85
Chapter 3.2.1 --- Cell lines --- p.85
Chapter 3.2.2 --- SYBR® Green PCR core kit --- p.85
Chapter 3.2.3 --- Chemicals --- p.85
Chapter 3.2.4 --- Solutions and buffers --- p.86
Chapter 3.2.5 --- Enzymes and nucleic acids --- p.86
Chapter 3.3 --- Methods --- p.88
Chapter 3.3.1 --- Preparation of total RNA from PGE2-induced JCS-14 and JCS cells --- p.88
Chapter 3.3.1.1 --- Preparation of cell lysates --- p.88
Chapter 3.3.1.2 --- Isolation of total RNA --- p.88
Chapter 3.3.2 --- Reverse transcription (RT) --- p.88
Chapter 3.3.3 --- Design of real-time PCR primers --- p.88
Chapter 3.3.4 --- Determination of relative efficiency of target and reference amplification by validation experiment --- p.89
Chapter 3.3.5 --- Confirmation of expression profile of identified genes in JCS-14 and JCS cells by comparative CT method in real-time PCR --- p.90
Chapter 3.4 --- Results --- p.91
Chapter 3.4.1 --- Analysis of total RNA from PGE2-induced JCS-14 and JCS cells --- p.91
Chapter 3.4.2 --- Validation experiment of real-time PCR primers --- p.93
Chapter 3.4.3 --- Expression profile of specific genes in JCS-14 and JCS cells by comparative CT method --- p.101
Chapter 3.5 --- Discussion --- p.114
Chapter 3.5.1 --- Study of gene expression profiles in JCS-14 and JCS cell differentiation --- p.114
Chapter 3.5.2 --- Transcription analysis by real-time PCR --- p.114
Chapter 3.5.3 --- Gene expression profiles during PGE2-induced JCS-14 and JCS cell differentiation --- p.115
Chapter Chapter Four --- Inhibition of specific gene expression in WEHI-3B JCS-14 and JCS cells using antisense blocking technique
Chapter 4.1 --- Introduction --- p.121
Chapter 4.1.1 --- Antisense technique --- p.122
Chapter 4.1.2 --- Design of antisense oligonucleotides --- p.125
Chapter 4.1.3 --- Transfer of oligonucleotides to cells --- p.128
Chapter 4.2 --- Materials --- p.129
Chapter 4.2.1 --- Cell lines --- p.129
Chapter 4.2.2 --- Chemicals --- p.129
Chapter 4.2.3 --- Reagents --- p.129
Chapter 4.2.4 --- Solutions --- p.129
Chapter 4.3 --- Methods --- p.131
Chapter 4.3.1 --- Design of antisense oligonucleotides --- p.131
Chapter 4.3.2 --- Transfection of oligonucleotides into cells --- p.134
Chapter 4.3.3 --- Morphological study of PGE2-induced JCS-14 and JCS cells --- p.134
Chapter 4.4 --- Results --- p.135
Chapter 4.4.1 --- Effect of antisense oligonucleotides on JCS-14 cell differentiation --- p.135
Chapter 4.4.2 --- Effect of antisense oligonucleotides on JCS cell differentiation --- p.136
Chapter 4.5 --- Discussion --- p.146
Chapter 4.5.1 --- Effects of antisense B-myb on JCS-14 and JCS cell differentiation --- p.146
Chapter 4.5.2 --- Effects of antisense thyroid hormone receptor (c-erbA) and transcription terminator factor (TTF-1) on JCS-14 and JCS cell differentiation --- p.147
Chapter Chapter Five --- General Discussion
Chapter 5.1 --- Introduction --- p.148
Chapter 5.2 --- Differentiation program triggered by Prostaglandin E2 --- p.148
Chapter 5.2.1 --- Lineage preference during differentiation --- p.148
Chapter 5.2.2 --- Differentially expressed genes during PGE2-induced JCS-14 cell differentiation --- p.149
Chapter 5.2.3 --- Expression patterns of the three differentially expressed genes in PGE2-induced JCS-14 and JCS cells --- p.149
Chapter 5.2.4 --- Antisense blocking during differentiation --- p.151
Chapter 5.3 --- Further studies --- p.152
References --- p.154
| Titel: |
Molecular study of differentially expressed genes in prostaglandin E₂ induced WEHI-3B JCS-14 and JCS cell differentiation.
|
|---|---|
| Autor/in / Beteiligte Person: | Chan, Sin-Man. ; Chinese University of Hong Kong Graduate School. Division of Biology. |
| Link: | |
| Veröffentlichung: | 2003 |
| Medientyp: | Nachschlagewerk |
| Schlagwort: |
|
| Sonstiges: |
|