Kong Ka-lai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 126-149). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Abstract (Chinese Version) --- p.vi / Table of Contents --- p.ix / List of Figures/Table --- p.xiii / List of Abbreviations --- p.xvi / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- An Overview on Cancer --- p.1 / Chapter 1.2 --- Biological Effects of Chlorophyllin --- p.7 / Chapter 1.2.1 --- CHL as Photosensitizer --- p.7 / Chapter 1.2.2 --- CHL as Antioxidant --- p.8 / Chapter 1.2.3 --- CHL as Anticarcinogenic Agent --- p.9 / Chapter 1.3 --- Regulation of Cell Cycle --- p.13 / Chapter 1.3.1 --- Cell-Cycle Checkpoints --- p.13 / Chapter 1.3.2 --- Cell-Cycle Regulatory Proteins --- p.15 / Chapter 1.4 --- Regulation of Mitogen-Activated Protein Kinase (MAPK) Signaling Cascade --- p.21 / Chapter 1.5 --- Programmed Cell Death (or Apoptosis) --- p.27 / Chapter 1.5.1 --- Regulation of Caspase-Dependent Apoptosis --- p.28 / Chapter 1.5.2 --- Regulation of Caspase-Independent Cell Death --- p.32 / Chapter 1.5.3 --- Bcl-2 Family Proteins in Modulation of Cell Death --- p.32 / Chapter 1.6 --- In Vivo Antitumor Screening System --- p.37 / Chapter 1.7 --- Aims of the Present Study --- p.38 / Chapter Chapter 2 --- In Vitro Studies of the Anticancer Effect of Chlorophyllin / Chapter 2.1 --- Introduction --- p.39 / Chapter 2.1.1 --- DNA-Flow Cytometric Analysis --- p.51 / Chapter 2.1.2 --- Western Blot Analysis --- p.54 / Chapter 2.2 --- Materials and Methods --- p.56 / Chapter 2.2.1 --- Maintenance of Cell Lines --- p.56 / Chapter 2.2.2 --- Cytotoxic and Cytostatic Effects on the Cancer Cells --- p.56 / Chapter 2.2.3 --- DNA-Flow Cytometric Analysis --- p.60 / Chapter 2.2.4 --- Western Blot Analysis --- p.61 / Chapter 2.2.5 --- JC-1 Mitochondrial Potential Sensor --- p.64 / Chapter 2.2.6 --- Caspase Inhibitors --- p.65 / Chapter 2.2.7 --- Statistical Analysis --- p.66 / Chapter 2.2.8 --- Densitometric Analysis --- p.66 / Chapter 2.3 --- Results --- p.67 / Chapter 2.3.1 --- Effects of CHL on the Growth of Human Cancer Cells by MTT Assay --- p.67 / Chapter 2.3.2 --- Effect of CHL on the Proliferation of MCF-7 Cells by Chemi-BrdU Incorporation --- p.69 / Chapter 2.3.3 --- Effect of CHL on Cell Cycle of MCF-7 Cells --- p.71 / Chapter 2.3.4 --- Effect of CHL on the Cyclin D1 Expression in MCF-7 Cells --- p.74 / Chapter 2.3.5 --- Effects of CHL on JNK and c-Jun Expressions and Their Phosphorylations in MCF-7 Cells --- p.76 / Chapter 2.3.6 --- Effect of CHL on DNA fragmentation in MCF-7 Cells --- p.78 / Chapter 2.3.7 --- Effect of CHL on Mitochondrial Membrane Potential of MCF-7 Cells --- p.80 / Chapter 2.3.8 --- Effects of CHL on the PARP Expression and Cleavage in MCF-7 Cells --- p.83 / Chapter 2.3.9 --- "Effects of CHL on Bcl-2, Bcl-xL and Bad Expressions in MCF-7 Cells" --- p.85 / Chapter 2.3.10 --- Effects of CHL on Caspase Activations in MCF-7 Cells --- p.88 / Chapter 2.3.11 --- Effects of Caspase Inhibitors on the CHL-Induced Apoptosis in MCF-7 Cells --- p.90 / Chapter 2.4 --- Discussion --- p.93 / Chapter Chapter 3 --- In Vivo Studies of the Anticancer Effect of Chlorophyllin / Chapter 3.1 --- Introduction --- p.104 / Chapter 3.2 --- Materials and Methods --- p.106 / Chapter 3.2.1 --- Transplantation of MCF-7 Cells into the Nude Mice and Treatment --- p.106 / Chapter 3.2.2 --- Western Blot Analysis --- p.107 / Chapter 3.2.3 --- Statistical Analysis --- p.107 / Chapter 3.3 --- Results --- p.108 / Chapter 3.3.1 --- In Vivo Antitumor Activity of CHL --- p.108 / Chapter 3.3.2 --- In Vivo Effects of CHL on Cyclin D1 and Bcl-2 Expressions in MCF-7 Solid Tumor --- p.111 / Chapter 3.4 --- Discussion --- p.113 / Chapter Chapter 4 --- General Discussion --- p.115 / References --- p.126

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Qin, Jing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 162-218). / Abstract also in Chinese. / Acknowledgment --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Table of Content --- p.vi / List of Figures --- p.x / List of Tables --- p.xv / Abbreviations --- p.xvii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Ganoderma spp --- p.1 / Chapter 1.1.1 --- Introduction of Ganoderma spp --- p.1 / Chapter 1.1.2 --- Bioactivities of Ganoderma spp --- p.3 / Chapter 1.1.3 --- Endocrine system and breast cancer --- p.11 / Chapter 1.1.3.1 --- Estrogen --- p.11 / Chapter 1.1.3.2 --- Estrogen receptors --- p.12 / Chapter 1.1.3.3 --- Estrogen responsive genes --- p.15 / Chapter 1.1.3.3.1 --- pS2 --- p.15 / Chapter 1.1.3.3.2 --- Progesterone receptor --- p.18 / Chapter 1.1.3.4 --- Androgen --- p.21 / Chapter 1.1.3.5 --- Androgen receptor --- p.23 / Chapter 1.1.3.6 --- Androgen responsive gene --- p.24 / Chapter 1.1.3.6.1 --- Transmembrane prostate androgen-induced RNA --- p.24 / Chapter 1.1.3.6.2 --- Uridine diphosphate glucose dehydrogenase --- p.26 / Chapter 1.1.3.7 --- Breast cancer --- p.26 / Chapter 1.2 --- "Endocrine Disruption of p,p '-DDE" --- p.28 / Chapter 1.2.1 --- Introduction of p´ةp '-DDE --- p.28 / Chapter 1.2.2 --- "p,p '-DDE in environments" --- p.29 / Chapter 1.2.3 --- "p,p '-DDE in human body" --- p.32 / Chapter 1.2.4 --- "p,p '-DDE and reproductive system" --- p.33 / Chapter 1.2.5 --- Endocrine disruptor --- p.35 / Chapter 1.2.6 --- "Action mechanism of p,p '-DDE on endocrine system" --- p.37 / Chapter 1.2.7 --- Apoptosis --- p.39 / Chapter 1.3 --- Food therapy against endocrine disruption --- p.41 / Chapter 1.3.1 --- Food therapy and functional food --- p.41 / Chapter 1.3.2 --- Ganoderma as a Functional food --- p.47 / Chapter 1.3.3 --- Cancer prevention by dietary agents --- p.47 / Chapter 1.3.4 --- Hormone therapy --- p.48 / Chapter 1.3.5 --- Hormone-related properties of Ganoderma spp --- p.50 / Chapter 1.4 --- The aim of the study --- p.51 / Chapter Chapter 2 --- Materials and Methods --- p.52 / Chapter 2.1 --- Ganoderma samples --- p.52 / Chapter 2.2 --- Artificial cultivation of Ganoderma spp --- p.54 / Chapter 2.3 --- Molecular identification of Ganoderma spp --- p.55 / Chapter 2.3.1 --- Extraction of genomic DNA --- p.55 / Chapter 2.3.2 --- Gene-specific polymerase chain reaction (PCR) --- p.56 / Chapter 2.3.3 --- Gel electrophoresis --- p.56 / Chapter 2.3.4 --- Purification of PCR amplified product for sequencing --- p.57 / Chapter 2.3.5 --- Cycle-sequencing --- p.57 / Chapter 2.3.6 --- Sequencing --- p.58 / Chapter 2.3.7 --- Sequence analysis --- p.58 / Chapter 2.4 --- Chemical analyses of Ganoderma spp --- p.59 / Chapter 2.4.1 --- Polysaccharide preparations --- p.59 / Chapter 2.4.2 --- Terpene profile --- p.60 / Chapter 2.4.3 --- Fatty acid profile --- p.60 / Chapter 2.5 --- Anti-oxidation activities --- p.61 / Chapter 2.5.1 --- Superoxide radical scavenging assay --- p.61 / Chapter 2.5.2 --- DPPH radical scavenging assay --- p.62 / Chapter 2.6 --- Anti-proliferation effect on human breast cancer cells --- p.62 / Chapter 2.7 --- Hormone-like effects --- p.63 / Chapter 2.7.1 --- E-screen test --- p.63 / Chapter 2.7.2 --- In vitro estrogen receptors (ERs) competitor binding assays --- p.64 / Chapter 2.7.3 --- "Recombinant yeast cell based ER-, AR- and PGR-responsible promoter assays" --- p.65 / Chapter 2.7.3.1 --- Recombinant yeasts --- p.65 / Chapter 2.7.3.2 --- Growth medium for recombinant yeasts --- p.66 / Chapter 2.7.3.3 --- "ER, AR and PGR assays" --- p.67 / Chapter 2.7.3.4 --- β-Galactosidase assay --- p.67 / Chapter 2.7.4 --- Real time PCR --- p.68 / Chapter 2.8 --- Flow cytometry --- p.71 / Chapter 2.9 --- Comet assay --- p.71 / Chapter 2.10 --- DNA microarray --- p.73 / Chapter 2.10.1 --- Total RNA isolation --- p.73 / Chapter 2.10.2 --- cDNA synthesis --- p.73 / Chapter 2.10.3 --- Preparation of labelled cDNA --- p.74 / Chapter 2.10.4 --- cDNA purification --- p.74 / Chapter 2.10.5 --- Oligo GEArray hybridization --- p.75 / Chapter 2.10.6 --- Chemiluminescent detection --- p.76 / Chapter 2.10.7 --- Data analysis --- p.77 / Chapter Chapter 3 --- Results --- p.78 / Chapter 3.1 --- Analysis of Ganderma spp --- p.78 / Chapter 3.1.1 --- Mycelia and fruiting bodies --- p.78 / Chapter 3.1.2 --- Identification of Ganoderma spp --- p.79 / Chapter 3.1.3 --- Chemical properties of samples --- p.80 / Chapter 3.1.4 --- Anti-oxidation activities --- p.90 / Chapter 3.1.5 --- Anti-proliferation effect on human breast cancer cells --- p.90 / Chapter 3.1.6 --- Hormone-like bioactivities --- p.93 / Chapter 3.1.6.1 --- E-screen test --- p.93 / Chapter 3.1.6.2 --- In vitro estrogen receptors (ERs) competitor binding assays --- p.94 / Chapter 3.1.6.3 --- "Recombinant yeast cell-based ER-, AR- and PGR-responsible promoter assays" --- p.95 / Chapter 3.1.6.4 --- ER- and AR-pathway gene expression by real time PCR --- p.97 / Chapter 3.2 --- "Action mechanism of p,p' -DDE" --- p.99 / Chapter 3.2.1 --- E-screen --- p.99 / Chapter 3.2.2 --- In vitro estrogen receptors (ERs) competitor binding assays --- p.101 / Chapter 3.2.3 --- Recombinant yeast cell based ER- and AR-responsible promoter assays --- p.103 / Chapter 3.2.4 --- ER- and AR-pathway gene expression by real time PCR --- p.106 / Chapter 3.3 --- Ganoderma tsugae mycelia extract against p.p' -DDE --- p.109 / Chapter 3.3.1 --- E-screen test --- p.109 / Chapter 3.3.2 --- ER- and AR-pathway gene expression by real time PCR --- p.110 / Chapter 3.3.3 --- Analysis of cell cycle --- p.112 / Chapter 3.3.4 --- Analysis of DNA damage --- p.114 / Chapter 3.3.5 --- Analysis of sub-G1 peak --- p.117 / Chapter 3.3.6 --- DNA damage and apoptosis relative gene expression by real time PCR --- p.120 / Chapter 3.3.7 --- DNA microarray --- p.121 / Chapter Chapter 4 --- Discussion --- p.131 / Chapter 4.1 --- Analysis of Ganoderma spp --- p.131 / Chapter 4.2 --- Effects of p.p´ة-DDE --- p.144 / Chapter 4.3 --- Protective effects of G. tsugae against p.p' -DDE --- p.151 / Chapter 4.4 --- Further investigation --- p.159 / Chapter 4.5 --- Conclusion --- p.160 / References --- p.162