Thesis (Ph.D.)--Baylor University, 2008. / In abstract the '50' in IC50 is subscript. Includes bibliographical references (p. 349-357).

Thesis (Ph. D.)--Michigan State University. Dept. of Biochemistry, 2007. / Title from PDF t.p. (viewed on Apr. 16, 2009) Includes bibliographical references. Also issued in print.

Thesis (Ph. D.)--University of Hong Kong, 2007. / Title proper from title frame. Also available in printed format.

Thesis (M.S.)--Villanova University, 2009. / Biology Dept. Includes bibliographical references.

published_or_final_version / abstract / Chemistry / Doctoral / Doctor of Philosophy

published_or_final_version / Dentistry / Doctoral / Doctor of Philosophy

Lai, Tsz Ching. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 121-136). / Abstracts in English and Chinese. / Acknowledgements / Abstract / 摘要 / Content / List of tables / List of figures / List of abbreviations / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Introduction of food market trends in Hong Kong and mushroom productivity in the world --- p.1 / Chapter 1.1.1 --- Agrocybe aegerita --- p.1 / Chapter 1.1.2 --- Pleurotus spp --- p.2 / Chapter 1.1.3 --- Pholiota nameko --- p.3 / Chapter 1.2 --- Objectives --- p.5 / Chapter Chapter 2: --- Chemical assays for in vitro antioxidative properties of mushroom extracts --- p.6 / Chapter 2.1 --- Introduction --- p.6 / Chapter 2.1.1 --- Reactive oxygen species (ROS) --- p.6 / Chapter 2.1.1.1 --- Definition of ROS --- p.6 / Chapter 2.1.1.2 --- Sources of ROS --- p.6 / Chapter 2.1.1.2.1 --- Endogenous sources of ROS --- p.6 / Chapter 2.1.1.2.2 --- Exogenous sources of ROS --- p.8 / Chapter 2.1.1.3 --- Damaging effects of ROS --- p.8 / Chapter 2.1.2 --- Antioxidants --- p.10 / Chapter 2.1.2.1 --- Mechanism of action --- p.10 / Chapter 2.1.2.2 --- Sources of antioxidants --- p.11 / Chapter 2.1.2.2.1 --- Dietary antioxidants --- p.11 / Chapter 2.1.2.2.2 --- Antioxidants in edible mushrooms --- p.12 / Chapter 2.1.2.2.3 --- Phenolic compounds in mushrooms --- p.13 / Chapter 2.2 --- Materials and Methods --- p.16 / Chapter 2.2.1 --- Materials --- p.16 / Chapter 2.2.1.1 --- Mushroom fruiting bodies --- p.16 / Chapter 2.2.2 --- Principles of Methods and Experimental Protocols --- p.17 / Chapter 2.2.2.1 --- Sample preparation --- p.17 / Chapter 2.2.2.2 --- Evaluation of antioxidant capacity --- p.18 / Chapter 2.2.2.2.1 --- DPPH radical scavenging activity --- p.18 / Chapter 2.2.2.2.2 --- Superoxide anion scavenging activity --- p.19 / Chapter 2.2.2.2.3 --- Hydroxyl radical scavenging activity --- p.20 / Chapter 2.2.2.2.4 --- Hydrogen peroxide scavenging activity --- p.22 / Chapter 2.2.2.3 --- Determination of phenolic compounds --- p.24 / Chapter 2.2.2.3.1 --- Total phenolic content --- p.24 / Chapter 2.2.2.3.2 --- Identification of phenolic acids --- p.25 / Chapter 2.2.3 --- Statistical analysis --- p.27 / Chapter 2.3 --- Results and Discussion --- p.28 / Chapter 2.3.1 --- Extraction yield --- p.28 / Chapter 2.3.2 --- Evaluation of antioxidant capacity --- p.29 / Chapter 2.3.2.1 --- DPPH radical scavenging activity --- p.29 / Chapter 2.3.2.2 --- Superoxide anion scavenging activity --- p.31 / Chapter 2.3.2.3 --- Hydroxyl radical scavenging activity --- p.33 / Chapter 2.3.2.4 --- Hydrogen peroxide scavenging activity --- p.35 / Chapter 2.3.2.5 --- Comparison of the effective concentrations (EC50) of mushroom water extracts in different antioxidant assays --- p.37 / Chapter 2.3.3 --- Determination of phenolic compounds --- p.38 / Chapter 2.3.3.1 --- Total phenolic content --- p.38 / Chapter 2.3.3.2 --- Identification of phenolic acids --- p.39 / Chapter 2.4 --- Summary --- p.45 / Chapter Chapter 3: --- Anti-angiogenic properties of the Aa water extract --- p.46 / Chapter 3.1 --- Introduction --- p.46 / Chapter 3.1.1 --- Angiogenesis --- p.46 / Chapter 3.1.1.1 --- Process of angiogenesis --- p.46 / Chapter 3.1.1.2 --- Regulations of angiogenesis --- p.47 / Chapter 3.1.1.2.1 --- Fibroblast growth factor (bFGF) --- p.47 / Chapter 3.1.1.2.2 --- Vascular endothelial growth factor (VEGF) --- p.48 / Chapter 3.1.2 --- Tumor angiogenesis --- p.49 / Chapter 3.1.2.1 --- ROS generation in tumor cells --- p.50 / Chapter 3.1.2.2 --- Hydrogen peroxide and VEGF --- p.51 / Chapter 3.1.2.3 --- Previous studies on tumor angiogenesis --- p.52 / Chapter 3.1.2.3.1 --- ROS and endothelial cells proliferation --- p.52 / Chapter 3.1.2.3.2 --- VEGF and endothelial cells functions --- p.53 / Chapter 3.1.3 --- Use of antioxidants in cancer treatment --- p.53 / Chapter 3.1.3.1 --- Antioxidant use of cancer therapy --- p.53 / Chapter 3.1.3.2 --- Antioxidant and endothelial cells functions --- p.54 / Chapter 3.1.3.3 --- Anti-angiogenic effects of polyphenols --- p.56 / Chapter 3.1.3.3.1 --- Phenolic acids --- p.56 / Chapter 3.1.3.3.2 --- Tea catechin --- p.57 / Chapter 3.1.3.3.3 --- Resveratrol --- p.57 / Chapter 3.1.3.3.4 --- Genistein --- p.58 / Chapter 3.2 --- Principles of Methods and Experimental Protocols --- p.60 / Chapter 3.2.1 --- Sample preparation --- p.60 / Chapter 3.2.2 --- Toxicity of the Aa water extract --- p.60 / Chapter 3.2.2.1 --- Limulus amebocyte lysate (LAL) test --- p.60 / Chapter 3.2.2.2 --- Toxicity towards normal cells --- p.61 / Chapter 3.2.2.2.1 --- Cell line and its subculture --- p.61 / Chapter 3.2.2.2.2 --- Colorimetric (MTT) assay --- p.62 / Chapter 3.2.3 --- Effect of the Aa water extract on cancer cells --- p.63 / Chapter 3.2.3.1 --- Cell line and its subculture --- p.63 / Chapter 3.2.3.2 --- Redox status --- p.63 / Chapter 3.2.3.3 --- VEGF secretion --- p.65 / Chapter 3.2.4 --- In vitro cell culture anti-angioenesis analysis --- p.66 / Chapter 3.2.4.1 --- Cell line and its subculture --- p.66 / Chapter 3.2.4.2 --- Endothelial cells proliferation --- p.67 / Chapter 3.2.4.3 --- Endothelial cells migration --- p.68 / Chapter 3.2.4.3.1 --- Wound healing assay --- p.68 / Chapter 3.2.4.3.2 --- Transwell culture insert assay --- p.69 / Chapter 3.2.4.4 --- Endothelial cells tubule formation --- p.71 / Chapter 3.2.5 --- In vitro organ culture anti-angiogenesis analysis --- p.72 / Chapter 3.2.5.1 --- Aortic ring assay --- p.72 / Chapter 3.2.6 --- Statistical analysis --- p.74 / Chapter 3.3 --- Results and Discussions --- p.75 / Chapter 3.3.1 --- Toxicity of the Aa water extract --- p.75 / Chapter 3.3.1.1 --- Limulus amebocyte lysate (LAL) test --- p.75 / Chapter 3.3.1.2 --- Toxicity towards normal cells --- p.75 / Chapter 3.3.2 --- Effect of the Aa water extract on cancer cells --- p.77 / Chapter 3.3.2.1 --- Redox status --- p.77 / Chapter 3.3.2.2 --- VEGF secretion --- p.79 / Chapter 3.3.2.3 --- Relationship between intracellular ROS and VEGF secretion detected --- p.80 / Chapter 3.3.3 --- Effect of the Aa water extract on angiogenesis --- p.82 / Chapter 3.3.3.1 --- Endothelial cells proliferation --- p.82 / Chapter 3.3.3.2 --- Endothelial cells migration --- p.84 / Chapter 3.3.3.2.1 --- Wound healing assay --- p.84 / Chapter 3.3.3.2.2 --- Transwell culture insert assay --- p.87 / Chapter 3.3.3.3 --- Endothelial cells tubule formation --- p.90 / Chapter 3.3.3.4 --- Aortic ring assay --- p.97 / Chapter 3.3.4 --- Effect of phenolic acids on endothelial cells --- p.101 / Chapter 3.3.4.1 --- Endothelial cells proliferation --- p.101 / Chapter 3.3.4.2 --- Endothelial cells migration --- p.102 / Chapter 3.3.4.2.1 --- Wound healing assay --- p.102 / Chapter 3.3.4.2.2 --- Transwell culture insert assay --- p.105 / Chapter 3.3.4.3 --- Endothelial cells tubule formation --- p.106 / Chapter 3.3.4.4 --- Aortic ring assay --- p.112 / Chapter 3.4 --- Summary --- p.116 / Chapter Chapter 4 --- Conclusions and future works --- p.118 / References --- p.121

Angiogenesis is one of the essential hallmarks of cancer, typically breast cancer. Signaling from VEGFR-2 is necessary for the execution of VEGF-induced proliferation, migration, and tube formation of cultured endothelial cells in vitro and the onset of angiogenesis on tumors in vivo. Ellagic acid is a naturally existing small molecular polyphenol widely found in fruits and vegetables. It was reported that ellagic aicd interfered with some angiogenesis-dependent pathologies. Yet the mechanisms involved were not fully understood. Thus we analyzed its anti-angiogenesis effects and mechanisms on human breast cancer utilizing in vitro and in vivo methodologies. Besides, the in silico analysis was carried out to further analyze the structure-based interaction between ellagic aicd and VEGFR-2. The influences of ellagic aicd on VEGF-induced endothelial cells were studied by proliferation, tube formation and migration in vitro experiments. Kinase activity assay and western blotting were utilized to explore the effects of ellagic aicd on VEGFR-2 induced signaling pathway. Organ-based chick aortic ring model, in vivo Chorioallantoic membrane model and in vivo breast cancer xenografts were built to determine the anti-angiogenesis effects of ellagic aicd. Besides, software LigandFit algorithm in Discovery Studio 2.1 (Accelrys Inc., San Diego, CA) was applied to further understand the structure-based interaction between ellagic aicd and VEGFR-2. We found that ellagic aicd impeded a series of VEGF-induced angiogenesis processes including proliferation, migration and tube formation of endothelial cells. Besides, it directly inhibited VEGFR-2 tyrosine kinase activity and its downstream signaling pathways including MAPK and PI3K/Akt on endothelial cells. Ellagic aicd also obviously inhibited sprouts formation from chicken aorta and neo-vessel formation in chick chorioallantoic membrane. The growth and the P-VEGFR2 expression in breast tumors treated with ellagic aicd were also significantly suppressed. In the molecular docking simulation experiment, the structure-based interaction of VEGFR-2 with ellagic acid was found to be stable conformation by hydrogen bonds within residues Lys866 and Glu883 as well as by π–π interactions within residue Phe1045 at ATP binding pocket of VEGFR-2 catalytic domain. Taken together, ellagic aicd could exert anti-angiogenesis effects via VEGFR-2 signaling pathway in breast cancer. / published_or_final_version / Chinese Medicine / Master / Master of Philosophy

Thesis (Ph. D.)--University of Hong Kong, 2008. / Also available in print.

Thesis (Ph. D.)--University of Hong Kong, 2008. / Also available online.