Photodynamic activity of a glucoconjugated Silicon(IV) phthalocyanine on human colon adenocarcinoma.

Chan, Man Hung.Thesis (M.Phil.)--Chinese University of Hong Kong, 2009.Includes bibliographical references (leaves 111-126).Abstract also in Chinese.Examination Committee List --- p.iiDeclaration --- p.iiiAcknowledgements --- p.iv摘要（Abstract in Chinese) --- p.viAbstract --- p.viiiList of Abbreviations --- p.xList of Figures and Tables --- p.xiiTable of Content --- p.xivChapter Chapter 1 --- Introduction --- p.1Chapter 1.1 --- Background of photodynamic therapy (PDT) --- p.2Chapter 1.1.1 --- History of PDT --- p.2Chapter 1.1.2 --- Photochemistry --- p.3Chapter 1.1.3 --- Principal stages of PDT --- p.5Chapter 1.1.4 --- Light sources of PDT --- p.6Chapter 1.2 --- Anti-tumor effect of PDT --- p.8Chapter 1.2.1 --- Mode of cell death --- p.8Chapter 1.2.2 --- PDT-induced anti-tumor immunity --- p.9Chapter 1.3 --- Clinical applications of PDT --- p.11Chapter 1.3.1 --- Photofrin® --- p.11Chapter 1.3.2 --- Clinical applications of PDT --- p.13Chapter 1.3.3 --- Challenges of PDT for clinical applications --- p.15Chapter 1.4 --- The development of new photosensitizers --- p.16Chapter 1.4.1 --- Targeted PDT --- p.16Chapter 1.4.2 --- Phthalocyanine --- p.18Chapter 1.5 --- Objective of my study --- p.21Chapter Chapter 2 --- Materials and Methods --- p.23Chapter 2.1 --- Synthesis of glucosylated silicon(IV) phthalocyanine (SiPcGlu) --- p.24Chapter 2.2 --- In vitro studies --- p.24Chapter 2.2.1 --- Cell line and culture conditions --- p.24Chapter 2.2.2 --- Photodynamic treatment --- p.25Chapter 2.2.3 --- Cell viability assay --- p.27Chapter 2.2.4 --- Light dose effect on the photocytotoxicity of SiPcGlu-PDT --- p.27Chapter 2.2.5 --- Determination of reactive oxygen species (ROS) production by SiPcGlu-PDT --- p.29Chapter 2.2.6 --- Effect of antioxidants on the photocytotoxicity of SiPcGlu-PDT --- p.29Chapter 2.2.7 --- Determination of ROS production after SiPcGlu-PDT --- p.30Chapter 2.2.8 --- Glucose competitive assay --- p.30Chapter 2.2.9 --- Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay --- p.30Chapter 2.2.10 --- DNA fragmentation analysis by gel electrophoresis --- p.31Chapter 2.2.11 --- Annexin-V & propidium iodide staining assay --- p.32Chapter 2.2.12 --- Subcellular localization studies --- p.33Chapter 2.2.13 --- Detection of mitochondrial superoxide production --- p.34Chapter 2.2.14 --- Assessment of mitochondrial membrane potential --- p.34Chapter 2.2.15 --- Caspase-3 activity assay --- p.35Chapter 2.2.16 --- "Western blot analyses for cytochrome c, caspase-3, PARP and glucose-regulated protein 78 (GRP78)" --- p.36Chapter 2.2.17 --- Ca2+ release from endoplasmic reticulum (ER) --- p.37Chapter 2.3 --- In vivo studies --- p.37Chapter 2.3.1 --- HT29 tumor-bearing nude mice model --- p.37Chapter 2.3.2 --- In vivo photodynamic treatment --- p.39Chapter 2.3.3 --- Biodistribution of SiPcGlu --- p.39Chapter 2.3.4 --- Assay for plasma enzyme activities --- p.40Chapter 2.4 --- Statistical analysis --- p.41Chapter Chapter 3 --- Results --- p.42Chapter 3.1 --- In vitro studies --- p.43Chapter 3.1.1 --- SiPcGlu-PDT induced cytotoxicity on HT29 cells --- p.43Chapter 3.1.2 --- Light dose effect on cytotoxicity by SiPcGlu-PDT --- p.46Chapter 3.1.3 --- SiPcGlu-PDT induced ROS production --- p.48Chapter 3.1.4 --- SiPcGlu-PDT induced cell death through Type I and II photoreactions --- p.48Chapter 3.1.5 --- ROS production after SiPcGlu-PDT --- p.51Chapter 3.1.6 --- Glucose competitive Assay --- p.55Chapter 3.1.7 --- SiPcGlu-PDT induced apoptosis in HT29 cells --- p.57Chapter 3.1.8 --- Subcellular localization of SiPcGlu --- p.61Chapter 3.1.9 --- SiPcGlu-PDT induced mitochondrial changes --- p.66Chapter 3.1.10 --- SiPcGlu-PDT induced caspase activation --- p.68Chapter 3.1.11 --- SiPcGlu-PDT increased expression of ER chaperone GRP78 --- p.72Chapter 3.1.12 --- SiPcGlu-PDT induced release of Ca2+ from ER --- p.72Chapter 3.2 --- In vivo studies --- p.75Chapter 3.2.1 --- In vivo photodynamic activities --- p.75Chapter 3.2.2 --- Tissue distribution of SiPcGlu --- p.77Chapter 3.2.3 --- Analysis of intrinsic toxicity --- p.77Chapter Chapter 4 --- Discussion --- p.80Chapter 4.1 --- Physical Properties of SiPcGlu --- p.81Chapter 4.2 --- In vitro studies --- p.82Chapter 4.2.1 --- SiPcGlu-PDT exhibits a high potency in killing HT29 cells --- p.82Chapter 4.2.2 --- ROS production is responsible for the cytotoxic effect of SiPcGlu-PDT --- p.83Chapter 4.2.3 --- SiPcGlu-PDT induced apoptosis in HT29 cells --- p.85Chapter 4.2.4 --- SiPcGlu is localized in various membranous organelles --- p.87Chapter 4.2.5 --- SiPcGlu-PDT induced mitochondria-mediated apoptosis --- p.89Chapter 4.2.6 --- SiPcGlu-PDT induced ER stress --- p.93Chapter 4.3 --- In vivo studies --- p.96Chapter 4.3.1 --- SiPcGlu failed to target to tumor tissues --- p.96Chapter 4.3.2 --- SiPcGlu-PDT induced retardation in tumor growth --- p.99Chapter 4.3.3 --- SiPcGlu is a safe photosensitizer for PDT --- p.101Chapter Chapter 5 --- Conclusion and Future Perspectives --- p.103Chapter 5.1 --- Conclusion --- p.104Chapter 5.2 --- Future Perspectives --- p.106Chapter 5.2.1 --- In vitro studies --- p.106Chapter 5.2.1.1 --- Lysosomal pathway to cell death --- p.106Chapter 5.2.2 --- In vivo studies --- p.107Chapter 5.2.2.1 --- Pharmacokinetic studies --- p.107Chapter 5.2.2.2 --- Eradication of HT29 tumor by repeated dose of SiPcGlu --- p.108Chapter 5.2.2.3 --- SiPcGlu-PDT-induced anti-tumor immunity --- p.108Chapter 5.2.2.4 --- Enhancement of tumor selectivity by conjugating with biomolecules --- p.109References --- p.11