Molecular analysis of BRAF and microsatellite analysis of chromosome 14q in astrocytic tumors.

Jazyk: Chinese<br />English
Rok vydání: 2004
Předmět:
Druh dokumentu: Bibliografie
Popis: Chan Ching Yin.
Thesis submitted in: October 2003.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2004.
Includes bibliographical references (leaves 197-221).
in English and Chinese.
Acknowledgement --- p.i
p.iii
in Chinese --- p.vi
List of abbreviations --- p.ix
List of tables --- p.xv
List of figures --- p.xvi
Contents --- p.xviii
Chapter 1. --- Introduction --- p.1
Chapter 1.1. --- What are astrocytic tumors? --- p.1
Chapter 1.1.1. --- Histological characteristics and classification --- p.2
Chapter 1.1.2. --- Epidemiology --- p.2
Chapter 1.1.3. --- Treatment and patient survival --- p.4
Chapter 1.2. --- "Cytogenetics, molecular genetics and epigenetics of astrocytic tumors" --- p.6
Chapter 1.2.1. --- Cytogenetics --- p.6
Chapter 1.2.2. --- Genetic imbalances --- p.7
Chapter 1.2.3. --- Tumor suppressor genes --- p.13
Chapter 1.2.4. --- Oncogenes --- p.22
Chapter 1.2.5. --- Primary and secondary GBMs --- p.26
Chapter 1.3. --- Major pathways involved in astrocytic tumorigenesis --- p.30
Chapter 1.3.1. --- Cell cycle dysregulation and suppression of apoptosis --- p.30
Chapter 1.3.2. --- Promotion of proliferation and survival --- p.33
Chapter 1.4. --- BRAF mutation in human cancers --- p.38
Chapter 1.5. --- Other CNS tumors included in the current study --- p.52
Chapter 2. --- Aims of study --- p.61
Chapter 3. --- Materials and methods --- p.64
Chapter 3.1. --- Clinical materials --- p.64
Chapter 3.2. --- Cell lines --- p.75
Chapter 3.3. --- Cell culture --- p.77
Chapter 3.4. --- DNA extraction --- p.78
Chapter 3.4.1. --- Pre-treatment of samples --- p.78
Chapter 3.4.2. --- Cell lysis and protein removal --- p.80
Chapter 3.4.3. --- Precipitation of DNA --- p.81
Chapter 3.4.4. --- Determination of DNA concentration --- p.81
Chapter 3.5. --- Mutation analysis of BRAF by cycle sequencing --- p.83
Chapter 3.5.1. --- Amplification of BRAF exons --- p.83
Chapter 3.5.2. --- Cycle sequencing and automated gel electrophoresis --- p.84
Chapter 3.6. --- Immunohistochemistry of B-Raf and GFAP --- p.87
Chapter 3.6.1. --- Pre-treatment of samples --- p.87
Chapter 3.6.2. --- Detection of B-Raf and GFAP antigens by ABC method --- p.88
Chapter 3.6.3. --- Controls --- p.90
Chapter 3.7. --- Quantification of EGFR gene dosage by TaqMan based real-time PCR --- p.91
Chapter 3.7.1. --- Preparation of gene constructs --- p.92
Chapter 3.7.2. --- Primers and TaqMan probes --- p.93
Chapter 3.7.3. --- Experimental condition and PCR program --- p.95
Chapter 3.7.4. --- DNA standards --- p.95
Chapter 3.7.5. --- Controls --- p.96
Chapter 3.7.6. --- Experimental layout --- p.96
Chapter 3.8. --- Microsatellite analysis of chromosome 14q in astrocytic tumors --- p.97
Chapter 4. --- Results --- p.101
Chapter 4.1. --- Mutation analysis of BRAF --- p.101
Chapter 4.2. --- Immunohistochemistry of B-Raf protein --- p.107
Chapter 4.3. --- Quantification of EGFR gene dosage --- p.117
Chapter 4.4. --- Correlation between EGFR dosage and BRAF mutation --- p.128
Chapter 4.5. --- Correlation between EGFR dosage and B-Raf expression --- p.129
Chapter 4.6. --- Microsatellite analysis of chromosome 14q in astrocytic tumors --- p.131
Chapter 5. --- Discussions --- p.149
Chapter 5.1. --- BRAF mutations as common events in human cancers --- p.149
Chapter 5.2. --- BRAF mutation in CNS tumor specimens --- p.150
Chapter 5.2.1. --- Tumorigenic effect of the V599E substitution --- p.153
Chapter 5.2.2. --- V599E B-Raf mutant activation independent of Ras activation --- p.155
Chapter 5.2.3. --- Autocrine stimulation of Ras signaling in V599E B-Raf mutant --- p.156
Chapter 5.3. --- BRAF expression in astrocytic tumors --- p.159
Chapter 5.4. --- Mutually exclusive pattern between EGFR amplification and BRAF expression --- p.161
Chapter 5.4.1. --- Similar effect of EGFR activation and B-Raf activation --- p.163
Chapter 5.4.2. --- Mutual effects between Ras/Raf/Mek/Erk and Akt signaling --- p.164
Chapter 5.5. --- Microsatellite analysis of chromosome 14q in human cancers --- p.167
Chapter 5.6. --- Microsatellite analysis of chromosome 14q in astrocytic tumors --- p.170
Chapter 5.6.1. --- Finer mapping of common regions of deletion --- p.170
Chapter 5.6.2. --- Genes within the common regions of deletion --- p.173
Chapter 5.6.3. --- Overlapping deletion regions in astrocytic and non-CNS tumors --- p.186
Chapter 6. --- Further studies --- p.190
Chapter 6.1. --- Role of BRAF alterations in astrocytic tumors --- p.190
Chapter 6.2. --- B-Raf expression in astrocytic tumors and correlation with EGFR overexpression --- p.193
Chapter 6.3. --- Microsatellite analysis of 14q in astrocytic tumors --- p.194
Chapter 7. --- Conclusions --- p.195
Chapter 8. --- References --- p.198
Databáze: Networked Digital Library of Theses & Dissertations
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