Cheung, Tsz Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 97-116). / Abstracts in English and Chinese. / Thesis/Assessment Committee Members --- p.ii / Declaration for Plagiarism and Copyright --- p.iii / Abstract --- p.iv / 摘要 --- p.vi / Acknowledgements --- p.viii / Table of Contents --- p.ix / List of Abbreviations --- p.xii / List of Figures --- p.xv / List of Tables --- p.xvi / Chapter CHAPTER TWO: --- Introduction / Chapter 1.1 --- Background Information / Chapter 1.1.1 --- Breast Cancer --- p.1 / Chapter 1.1.2 --- General Statistics of Breast Cancer Worldwide and in Hong Kong --- p.1 / Chapter 1.1.3 --- Risk Factors for Breast Cancer --- p.2 / Chapter 1.1.4 --- Breast Cancer Treatment and Side Effects --- p.2 / Chapter 1.1.5 --- Types of Breast Cancer --- p.3 / Chapter 1.2 --- Estrogen and Estrogen Receptor / Chapter 1.2.1 --- Estrogen --- p.4 / Chapter 1.2.2 --- Estrogen Receptor --- p.5 / Chapter 1.2.3 --- Estrogen Receptor mediated Gene Transcription --- p.5 / Chapter 1.2.4 --- Estrogen Receptor Alpha and Estrogen Receptor Beta --- p.6 / Chapter 1.2.5 --- Estrogen Receptor Positive Breast Cancer and Treatment --- p.7 / Chapter 1.3 --- Estrogen metabolism and Cytochrome P450 family 1 (CYP1) members / Chapter 1.3.1 --- Estrogen Metabolism in Human --- p.9 / Chapter 1.3.2 --- CYP1A1 and CYP1B1 --- p.9 / Chapter 1.3.3 --- Estrogen Metabolism in Breast --- p.10 / Chapter 1.3.4 --- Carcinogenesis of Estrogens and Estrogen Metabolites --- p.13 / Chapter 1.3.5 --- The Importance of CYP1B1 in Carcinogenesis --- p.15 / Chapter 1.4 --- Aryl Hydrocarbon Receptor / Chapter 1.4.1 --- General Information of Aryl Hydrocarbon Receptor --- p.16 / Chapter 1.4.2 --- Signaling/Regulation Pathways of Aryl Hydrocarbon Receptor --- p.17 / Chapter 1.4.3 --- Crosstalk with Estrogen Receptor --- p.17 / Chapter 1.5 --- Introduction of Herba Scutellaria Barbata and its active ingredient Pheophorbide a --- p.19 / Chapter 1.6 --- Hyposthesis and Objectives --- p.21 / Chapter CHAPTER TWO: --- Direct Cytotoxic/Cytostatic Effect of Pheophorbide a / Chapter 2.1 --- Backgrounds --- p.22 / Chapter 2.2 --- Materials / Chapter 2.2.1 --- Chemicals --- p.24 / Chapter 2.2.2 --- Cell Lines --- p.26 / Chapter 2.2.3 --- "Cell Culture Mediums, Buffers and Consumables" / Chapter 2.2.3.1 --- Roswell Park Memorial Institute Tissue Culture Medium1640 (RPMI1640) --- p.26 / Chapter 2.2.3.2 --- RPMI 1640 (Phenol Red-free) --- p.26 / Chapter 2.2.3.3 --- Serum supplement - Fetal Bovine Serum (FBS) --- p.27 / Chapter 2.2.3.4 --- Serum supplement - Charcoal/Dextran Stripped FBS --- p.27 / Chapter 2.2.3.5 --- Antibiotics - Penicillin-Streptomycin (P/S) --- p.27 / Chapter 2.2.3.6 --- Trypsin (0.25%) with EDTA --- p.27 / Chapter 2.2.3.7 --- Trypsin (2.5%) (Phenol Red-free) with EDTA --- p.28 / Chapter 2.2.3.8 --- Dulbeccóةs Phosphate-Buffered Saline (D-PBS) --- p.28 / Chapter 2.2.3.9 --- Tissue Culture Flasks and Multi-well Plate --- p.28 / Chapter 2.2.3.10 --- Trypan Blue Solution --- p.29 / Chapter 2.2.4 --- Reagents for Direct Cytotoxity Test / Chapter 2.2.4.1 --- MTT Assay --- p.29 / Chapter 2.2.4.2 --- Tritiated Thymidine Incorporation Assay --- p.29 / Chapter 2.3 --- Methods / Chapter 2.3.1 --- Cell Culture --- p.30 / Chapter 2.3.2 --- Direct Cytotoxicity/Cytostatic Test / Chapter 2.3.2.1 --- MTT Assay --- p.31 / Chapter 2.3.2.2 --- Tritiated Thymidine Incorporation Assay --- p.32 / Chapter 2.3.3 --- Statistical Analysis --- p.32 / Chapter 2.4 --- Results / Chapter 2.4.1 --- The Cytotoxic Effect of Pheophorbide a --- p.34 / Chapter 2.4.2 --- The Combine Effect of Pheophorbide a with 17-β Estradiol and Tamoxifen Citrate --- p.34 / Chapter 2.5 --- Discussions --- p.48 / Chapter CHAPTER THREE: --- Mechanistic Study of Pheophorbide a / Chapter 3.1 --- Backgrounds --- p.53 / Chapter 3.2 --- Materials / Chapter 3.2.1 --- Real time PCR / Chapter 3.2.1.1 --- General Chemicals and Equipments --- p.54 / Chapter 3.2.1.2 --- RNA isolation --- p.55 / Chapter 3.2.1.3 --- Reverse Transcription --- p.55 / Chapter 3.2.1.4 --- Real Time PCR --- p.56 / Chapter 3.2.2 --- Western Blotting / Chapter 3.2.2.1 --- Microsome Isolation --- p.58 / Chapter 3.2.2.2 --- Measurement of Protein Concentration --- p.58 / Chapter 3.2.2.3 --- Western Blotting --- p.58 / Chapter 3.2.3 --- Estrogen Metabolism Assay / Chapter 3.2.3.1 --- Chemicals --- p.59 / Chapter 3.2.3.2 --- Estrogen Metabolites Extraction --- p.60 / Chapter 3.2.3.3 --- Liquid Chromatography/Mass Spectrometry --- p.60 / Chapter 3.3 --- Methods / Chapter 3.3.1 --- Real time PCR / Chapter 3.3.1.1 --- Cell Culture --- p.61 / Chapter 3.3.1.2 --- RNA Isolation and Reverse Transcription --- p.61 / Chapter 3.3.1.3 --- Real Time PCR --- p.62 / Chapter 3.3.2 --- Western Blotting / Chapter 3.3.2.1 --- Cell Culture --- p.63 / Chapter 3.3.2.2 --- Microsome Isolation --- p.63 / Chapter 3.3.2.3 --- Measurement of Protein Concentration --- p.64 / Chapter 3.3.2.4 --- Western Blotting --- p.64 / Chapter 3.3.3 --- Estrogen Metabolism Assay / Chapter 3.3.3.1 --- Preparation of Calibration Standard --- p.65 / Chapter 3.3.3.2 --- Cell Culture --- p.66 / Chapter 3.3.3.3 --- Estrogen Metabolites Extraction --- p.66 / Chapter 3.3.3.4 --- Liquid Chromatography/Mass Spectrometry --- p.67 / Chapter 3.3.4 --- Statistical Analysis --- p.68 / Chapter 3.4 --- Results --- p.69 / Chapter 3.5 --- Discussions --- p.80 / Chapter CHAPTER FOUR: --- Overall Conclusion and Future Directions / Chapter 4.1 --- Significance of the Study --- p.87 / Chapter 4.2 --- Overall Conclusion --- p.87 / Chapter 4.3 --- Limitation and Difficulties of the Study --- p.89 / Chapter 4.4 --- Future Directions --- p.89 / Appendices / "Appendix I The Melting Curve of real time PCR for β-actin, CYP1A1 and CYP1B1" --- p.92 / Appendix II The Calibration Curve of BSA for Protein Concentration Measurement --- p.93 / Appendix III The Representative Peak of Estradiol Metabolite Standards with corresponding Retention Time --- p.94 / Appendix IV The Calibration Curve of Different Estrogen Metabolites for LC/MS --- p.95 / Appendix V The Accuracy and Precision of Quality Control of Estradiol Metabolites --- p.96 / Bibliography --- p.97
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326791 |
| Date | January 2009 |
| Contributors | Cheung, Tsz Yan., Chinese University of Hong Kong Graduate School. Division of Pharmacy. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xvi, 116 leaves : ill. (some col.) ; 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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