by Ng Mei Yu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 173-183). / Abstracts in English and Chinese. / Acknowledgements --- p.2 / Abbreviations --- p.3 / Abstracts --- p.4 / List of figures and tables --- p.7 / Contents Page No / General Introduction --- p.13 / Chapter CHAPTER ONE --- Biological Characterization of A431 Cells & SiHa Cells Under Different Microenvironments --- p.16 / Chapter 1.1 --- Introduction --- p.17 / Chapter 1.1.1 --- Microenvironment Surrounding Tumor Cells --- p.18 / Chapter 1.1.2 --- Hypoxic Environment --- p.20 / Chapter 1.1.2.1 --- The Hypoxic Chamber --- p.20 / Chapter 1.1.3 --- Reoxygenation --- p.22 / Chapter 1.1.4 --- Acidic Environment --- p.23 / Chapter 1.1.5 --- Glucose Depletion --- p.24 / Chapter 1.1.6 --- Irradiation --- p.25 / Chapter 1.2 --- Objectives --- p.26 / Chapter 1.3 --- Materials and Methods --- p.27 / Chapter 1.3.1 --- Materials --- p.27 / Chapter 1.3.2 --- Methods --- p.29 / Chapter 1.3.2.1 --- Cell Lines --- p.29 / Chapter 1.3.2.2 --- The Working Procedure for the Hypoxic Chamber --- p.30 / Chapter 1.3.2.3 --- "Aerobic, Hypoxic and Reoxygenated Conditions" --- p.33 / Chapter 1.3.2.4 --- Acidic Condition --- p.35 / Chapter 1.3.2.5 --- Glucose Depleted Condition --- p.36 / Chapter 1.3.2.6 --- Gamma-Irradiation --- p.37 / Chapter 1.3.2.7 --- Analysis of the Growth Pattern by MTT Assay and Cell Counting --- p.38 / Chapter 1.3.2.8 --- Cell Cycle Analysis --- p.39 / Chapter 1.3.2.9 --- Western Blot Analysis --- p.40 / Chapter 1.3.2.10 --- DNA Fragmentation Analysis --- p.42 / Chapter 1.4 --- Results --- p.43 / Chapter 1.4.1 --- Cell Proliferation Profile by MTT Assay --- p.43 / Chapter 1.4.1.1 --- Proliferation of cells under hypoxia --- p.43 / Chapter 1.4.1.2 --- Proliferation of cells under acidic pH environments --- p.49 / Chapter 1.4.1.3 --- Proliferation of cells under glucose depleted environment --- p.52 / Chapter 1.4.2 --- Distribution of cell cycles under different micro environments --- p.54 / Chapter 1.4.3 --- General Protein Expression Pattern by Western Blot Analysis --- p.57 / Chapter 1.4.4 --- Detection of Apoptosis by DNA Fragmentation Assay --- p.59 / Chapter 1.5 --- Discussion --- p.58 / Chapter CHAPTER TWO --- REACTION KINETICS OF A431 CELLS AND SiHa CELLS INDUCED BY EGF --- p.71 / Chapter 2.1 --- Introduction --- p.72 / Chapter 2.1.1 --- Structure of EGF and EGFR --- p.74 / Chapter 2.1.2 --- EGF Signaling Pathway --- p.76 / Chapter 2.2 --- Objectives --- p.79 / Chapter 2.3 --- Materials and Methods --- p.80 / Chapter 2.3.1 --- Materials --- p.80 / Chapter 2.3.2 --- Methods --- p.82 / Chapter 2.3.2.1 --- Cell Lines --- p.82 / Chapter 2.3.2.2 --- EGF Sensitivity Assay --- p.83 / Chapter 2.3.2.3 --- Combination Effect of Hypoxia and EGF --- p.83 / Chapter 2.3.2.4 --- Early Kinetics Analysis by Low EGF Concentration Treatment --- p.84 / Chapter 2.3.2.5 --- Late Kinetics Analysis by High EGF Concentration Treatment --- p.85 / Chapter 2.4 --- Results --- p.86 / Chapter 2.4.1 --- Sensitivity of A431 cells and SiHa cells to EGF by MTT Assay --- p.86 / Chapter 2.4.2 --- Early/Late Kinetics of EGF induced protein tyrosine phosphorylation Pattern --- p.90 / Chapter 2.4.3 --- Raf protein expression --- p.96 / Chapter 2.4.4 --- EGFR expression level --- p.100 / Chapter 2.5 --- Discussions --- p.104 / Chapter CHAPTER THREE --- IDENTIFICATION OF DIFFERENTIALLY EXPRESSED GENES IN A431 CELLS BY DIFFERENTIAL DISPLAY UNDER DIFFERENT TUMOR MICROENVIRONMENTS --- p.107 / Chapter 3.1 --- Introduction --- p.108 / Chapter 3.2 --- Materials and Methods --- p.112 / Chapter 3.2.1 --- Materials --- p.112 / Chapter 3.2.2 --- Methods --- p.114 / Chapter 3.2.2.1 --- Spheroid Cells --- p.114 / Chapter 3.2.2.2 --- Identification of Differentally Expressed Genes by RT-PCR --- p.117 / Chapter 3.2.2.3 --- Ligation and Cloning of Differentially Expressed cDNA --- p.120 / Chapter 3.2.2.4 --- Screening and Sequencing of the cDNA Inserts --- p.121 / Chapter 3.2.2.5 --- Northern Blot Analysis --- p.123 / Chapter 3.3 --- Results --- p.124 / Chapter 3.4 --- Discussions --- p.161 / GENERAL CONCLUSION --- p.165 / REFERENCES --- p.167
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_323848 |
| Date | January 2002 |
| Contributors | Ng, Mei Yu., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, 183 leaves : ill. ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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