Neuroblastoma is one of the most common solid tumors in patients below the age of 15. In this PhD project, the possible anti-tumor effects of the soy isoflavones on neuroblastoma cells have been investigated. A number of aspects of neuroblastoma cancer cell biology, including cell proliferation, cell cycle regulation, cell differentiation and apoptosis, intracellular signaling mechanisms, tumor invasiveness and metastatic properties, have been examined. Furthermore, novel antitumor properties of the soy isoflavones on neuroblastoma have also been quested for, hoping that through this PhD project, a better understanding of the potential anti-tumor effects of soy isoflavones on neuroblastoma cells can be obtained. In Chapter Three, we have demonstrated that the major soy isoflavones daidzein and genistein exerted potent anti-proliferative effect on murine neuroblastoma Neuro-2a (BU-1) cells. These two compounds were shown to modulate cell cycle distribution in BU-1 cells in different ways, possible through their differential effects on the expression of cell cycle regulatory proteins such as cyclins and cyclin-dependent kinase inhibitors. The anti-tumor effect of daidzein was found to be fairly specific to tumor cells, as daidzein only exhibited little, if any, direct cytotoxicity to normal murine cells such as bone marrow cells, macrophages, and thymocytes. Furthermore, the anti-proliferative effect of daidzein was unlikely to be attributed to its direct cytotoxicity to the BU-1 cells, but might be coupled with its ability to trigger the apoptosis and differentiation in the neuroblastoma cells. Our results show that daidzein could induce DNA fragmentation, ultrastructural changes, and the enrichment of cytoplasmic mono- and oligo-nucleosomes in the treated-BU-1 cells. Moreover, daidzein was shown to induce neuronal differentiation in the BU-1 cells, as indicated by morphological changes, and increased expression of the neuronal differentiation marker microtubule-associated protein-2, and acetylcholine esterase activity. In addition, we have also demonstrated that the signals of daidzein might be mediated by the estrogen receptor and nuclear factor-kappa B pathways. In Chapter Four, daidzein was shown to exert a differential an proliferative effect on three human neuroblastoma cell lines, including SK-N-DZ and SH-SY5Y. The most sensitive cell line, LA-N-1, was chosen for further mechanistic studies. It was found that daidzein-induced growth-inhibitory effect was coupled with the induction of neurite outgrowth and altered mRNA expression of the N-myc-related transcription factors. Moreover, daidzein was observed to modulate the invasiveness and metastatic properties of LA-N-1 cells, as indicated by the reduction of colony-forming ability, cell migratory ability, in vivo tumorigenicity, tumor vascularity, and angiogenic factors expression. Our results clearly suggest that the major soy isoflavone daidzein can exert its pleiotropic anti-tumor effects on both marine and human neuroblastoma cells. In Chapter Five, we isolated two stable actively proliferating subclones from the human neuroblastoma SH-SY5Y cells. We compared the sensitivities of the parental SH-SY5Y cells and the active subclones to the growth inhibition exerted by a number of conventional cancer chemotherapeutic agents and the soy isoflavone derivatives. We found that the major soy isoflavones daidzein and genistein were rather selective to the active subclones and this phenomenon was not observed with other chemotherapeutic agents. The anti-tumor action mechanisms of genistein on the most active subclone, designated as SH-SY5Y cl.6 cells, were examined in detail. It was found that genistein could induce apoptosis in SH-SY5Y cl.6 cells, as indicated by the induction of ultrastructural changes, phosphatidylserine externalization, and cytoplasmic enrichment of mono- and oligo-nucleosomes. The genistein-induced apoptosis in SH-SY5Y cl.6 cells was found to be both mitochondria and caspases-dependent, as mitochondrial membrane depolarization, cytosolic release of apoptotic mitochondrial factors and activation of caspase-3 were observed. To sum up, our results show that the major soy isoflavones, particularly daidzein and genistein, exhibit pleiotropic anti-tumor effects on both murine and human neuroblastoma cells, and they are also selective to the actively proliferating human neuroblastoma cells. (Abstract shortened by UMI.) / Lo, Fai-Hang. / "September 2007." / Adviser: Leung Kwok-Nam. / Source: Dissertation Abstracts International, Volume: 69-02, Section: B, page: 0949. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 255-287). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract in English and Chinese. / School code: 1307.

Guan, Lei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 123-145). / Abstracts in English and Chinese. / ACKNOWLEDGEMENT --- p.I / ABSTRACT --- p.II / LIST OF ABBREVIATIONS --- p.VII / TABLE OF CONTENT --- p.IIX / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Distribution and Origins --- p.1 / Chapter 1.3 --- History of Use --- p.2 / Chapter 1.4 --- Chemical Structure --- p.3 / Chapter 1.5 --- Physiologic Properties --- p.5 / Chapter 1.6 --- Absorption and Metabolism --- p.7 / Chapter 1.6.1 --- Concentration of Isoflavones in Plasma --- p.10 / Chapter 1.6.2 --- Urinary Excretion --- p.10 / Chapter 1.7 --- Healthy Effects --- p.11 / Chapter 1.7.1 --- Menopausal Symptom --- p.11 / Chapter 1.7.2 --- Cardiovascular Disease --- p.12 / Chapter 1.7.3 --- Osteoporosis --- p.13 / Chapter 1.7.4 --- Tumors --- p.14 / Chapter 1.7.4.1 --- Breast Cancer --- p.14 / Chapter 1.7.4.2 --- Prostate Cancer --- p.15 / Chapter 1.7.5 --- Alcohol Addiction --- p.16 / Chapter 1.7.6 --- Potential Adverse Effects --- p.16 / Chapter 1.8 --- Summary --- p.17 / Chapter Chapter 2 --- Determination of Isoflavones in Soybean and Gegen --- p.19 / Chapter 2.1 --- Introduction --- p.19 / Chapter 2.1.1 --- Classification and Structure of Phytoestrogens --- p.19 / Chapter 2.1.2 --- Isoflavones in Soybeans and Gegen --- p.21 / Chapter 2.1.3 --- Methods of Determination --- p.26 / Chapter 2.1.3.1 --- Isolation and Purification of Isoflavones --- p.26 / Chapter 2.1.3.2 --- Analytical Methods for Isoflavones in Soybeans and Gegen --- p.26 / Chapter 2.2 --- Objective --- p.28 / Chapter 2.3 --- Materials and Methods --- p.29 / Chapter 2.3.1 --- Extraction and Isolation of Soybean and Gegen Isoflavone Extracts --- p.29 / Chapter 2.3.2 --- HPLC Analysis --- p.29 / Chapter 2.3.2.1 --- Sample Preparation for the HPLC Analysis --- p.29 / Chapter 2.3.2.2 --- HPLC Analysis --- p.30 / Chapter 2.3.3.3 --- Qualitative Analysis of the Isoflavones and Their Glycosides in Soybean and Gegen --- p.30 / Chapter 2.4 --- Results --- p.31 / Chapter 2.4.1 --- Isoflavone Identification of Soybean Extract --- p.31 / Chapter 2.4.2 --- Isoflavone Identification of Gegen Extract --- p.33 / Chapter 2.5 --- Discussion --- p.35 / Chapter Chapter 3 --- Hypocholesterolemic Effects of Soybean and Gegen Isoflavone Extracts in Ovariectomized，Intact Male and Castrated Golden Syrian Hamsters --- p.38 / Chapter 3.1 --- Introduction --- p.38 / Chapter 3.2 --- Objective --- p.41 / Chapter 3.3 --- Materials and Methods --- p.42 / Chapter 3.3.1 --- Preparation of Soybean and Gegen Isoflavone Extracts --- p.42 / Chapter 3.3.2 --- Animals and Diets --- p.42 / Chapter 3.3.3 --- Serum Lipid and Lipoprotein Determinations --- p.45 / Chapter 3.3.4 --- Determination of Cholesterol Concentration in the Organs --- p.45 / Chapter 3.3.5 --- Statistics --- p.46 / Chapter 3.4 --- Results --- p.48 / Chapter 3.4.1 --- Food Intake and Body and Relative Organ Weights --- p.48 / Chapter 3.4.2 --- Effects of Soybean and Gegen Isoflavone Extracts on Serum and Organ Cholesterol in Ovariectomized and Intact Male and Castrated Hamsters --- p.56 / Chapter 3.5 --- Discussion --- p.61 / Chapter Chapter 4 --- Possible Developmental and Reproductive Toxicity of Soybean Isoflavones on SD Rats --- p.67 / Chapter 4.1 --- Introduction --- p.67 / Chapter 4.2 --- Objective --- p.70 / Chapter 4.3 --- Materials and Methods --- p.71 / Chapter 4.3.1 --- Diet --- p.71 / Chapter 4.3.2 --- Animals --- p.73 / Chapter 4.3.3 --- Study Design --- p.73 / Chapter 4.3.4 --- Measurement of Reproductive Hormones --- p.74 / Chapter 4.3.5 --- Measurement of Sperm Number --- p.74 / Chapter 4.3.6 --- Statistics --- p.75 / Chapter 4.4 --- Results --- p.77 / Chapter 4.4.1. --- Food Intake and Food Efficiency Ratio --- p.77 / Chapter 4.4.2 --- Growth Trend --- p.79 / Chapter 4.4.3 --- Organ Weight --- p.82 / Chapter 4.4.3.1 --- Absolute Organ Weight --- p.82 / Chapter 4.4.3.2 --- Relative Organ Weight --- p.84 / Chapter 4.4.4 --- Reproductive Hormone Levels --- p.86 / Chapter 4.4.5 --- Epididymis Parameters of Male Rats --- p.88 / Chapter 4.5 --- Discussion --- p.90 / Chapter Chapter 5 --- Possible Developmental and Reproductive Toxicity of Gegen Isoflavones on SD Rats --- p.97 / Chapter 5.1 --- Introduction --- p.97 / Chapter 5.2 --- Objective --- p.99 / Chapter 5.3 --- Materials and Methods --- p.100 / Chapter 5.3.1 --- Animals and Diets --- p.100 / Chapter 5.3.2 --- Study Design --- p.100 / Chapter 5.3.3 --- Statistics --- p.101 / Chapter 5.4 --- Results --- p.103 / Chapter 5.4.1 --- Food Consumption and Food Efficiency Ratio --- p.103 / Chapter 5.4.2 --- Growth Trend --- p.105 / Chapter 5.4.3 --- Organ Weights --- p.108 / Chapter 5.4.3.1 --- Absolute Organ Weights --- p.108 / Chapter 5.4.3.2 --- Relative Organ Weight --- p.110 / Chapter 5.4.4 --- Reproductive Hormone Levels --- p.112 / Chapter 5.4.5 --- Epididymis Parameters of Male Rats --- p.114 / Chapter 5.5 --- Discussion --- p.116 / Chapter Chapter 6 --- Conclusions --- p.121 / References --- p.123

Han, Li. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 132-140). / Abstract and appendixes also in Chinese. / Chapter 1.1 --- BACKGROUND & SIGNIFICANCE OF THE STUDY --- p.1 / Chapter 1.2 --- STRUCTURE OF THE THESIS --- p.4 / Chapter 2.1 --- BPH --- p.6 / Chapter 2.1.1 --- PREVALENCE OF BPH --- p.6 / Chapter 2.1.2 --- IMPACT OF BPH SYMPTOMS ON PATIENTS --- p.9 / Chapter 2.1.2.1 --- CLINICAL SYMPTOMS OF BPH --- p.9 / Chapter 2.1.2.2 --- IMPACT OF CLINICAL SYMPTOMS ON QUALITY OF LIFE --- p.10 / Chapter 2.1.3 --- IMPACT OF BPH MEDICAL MANAGEMENT ON PATIENTS --- p.11 / Chapter 2.1.3.1 --- MEDICAL MANAGEMENT OF BPH --- p.11 / Chapter 2.1.3.2 --- SIDE EFFECTS OF PHARMACOLOGICAL AND SURGICAL THERAPIES --- p.15 / Chapter 2.1.4 --- USE OF COMPLEMENTARY AND ALTERNATIVE THERAPY AMONG BPH PATIENTS --- p.19 / Chapter 2.1.4.1 --- PREVALENCE --- p.19 / Chapter 2.1.4.2 --- REASONS FOR TURNING TO CAM --- p.20 / Chapter 2.2 --- ISOFLAVONES --- p.43 / Chapter 2.2.1 --- ISOFLAVONES FUNCTION --- p.43 / Chapter 2.2.1.1 --- STRUCTURE --- p.43 / Chapter 2.2.1.2 --- FOOD SOURCES --- p.45 / Chapter 2.2.2 --- APPLICATION OF ISOFLAVONES IN BPH --- p.46 / Chapter 2.2.2.1 --- DOCUMENTED MECHANISM OF BPH --- p.46 / Chapter 2.2.2.2 --- STUDIES IN VITRO --- p.47 / Chapter 2.2.2.3 --- STUDIES IN VIVO --- p.48 / Chapter 2.2.2.4 --- EPIDEMIOLOGIC EVIDENCE --- p.49 / Chapter 2.3. --- RESEARCH GAP IN HUMAN STUDY --- p.50 / Chapter 3.1 --- STUDY DESIGN --- p.51 / Chapter 3.2 --- AIM --- p.51 / Chapter 3.3 --- STUDY POPULATION --- p.52 / Chapter 3.3.1 --- INCLUSION CRITERIA --- p.52 / Chapter 3.3.2 --- EXCLUSION CRITERIA --- p.52 / Chapter 3.3.3 --- SAMPLE SIZE ESTIMATION --- p.53 / Chapter 3.4 --- RANDOMIZATION --- p.54 / Chapter 3.4.1 --- GENERATION THE RANDOM ALLOCATION SEQUENCE AND DETAILS OF RESTRICTION OF RANDOMIZATION --- p.54 / Chapter 3.4.2 --- IMPLEMENTATION OF RANDOMIZATION --- p.54 / Chapter 3.5 --- BLINDING --- p.55 / Chapter 3.5.1 --- WHO WERE BLINDED --- p.55 / Chapter 3.6 --- INTERVENTION --- p.55 / Chapter 3.6.1 --- STUDY MEDICATIONS AND DOSAGE --- p.55 / Chapter 3.6.2 --- STUDY REGIMEN --- p.56 / Chapter 3.7 --- DATA COLLECTION --- p.56 / Chapter 3.8 --- OUTCOME MEASUREMENTS --- p.58 / Chapter 3.8.1 --- PRIMARY OUTCOME --- p.58 / Chapter 3.8.1.1 --- UROFLOWMETRY： PEAK URINE FLOW RATE (QMAX) --- p.58 / Chapter 3.8.2 --- SECONDARY OUTCOMES --- p.59 / Chapter 3.8.2.1 --- BLADDER SCAN： POST-VOIDING RESIDUAL VOLUME (PVR) --- p.59 / Chapter 3.8.2.2 --- SYMPTOMS SCORE (IPSS) --- p.60 / Chapter 3.8.2.3 --- QUALIFY OF LIFE --- p.61 / Chapter 3.8.2.4 --- SERUM PSA LEVEL --- p.62 / Chapter 3.8.2.5 --- URINALYSIS TEST --- p.62 / Chapter 3.8.3 --- AE/SAE --- p.63 / Chapter 3.8.3.1 --- SELF REPORTED AE/SAE --- p.63 / Chapter 3.8.3.2 --- SEXUAL HORMONE LEVEL --- p.64 / Chapter 3.8.3.3 --- SEXUAL RELATED QUALITY OF LIFE --- p.64 / Chapter 3.9 --- STATISTICAL ANALYSIS --- p.65 / Chapter 3.9.1 --- DESCRIPTIVE ANALYSIS --- p.65 / Chapter 3.9.2 --- COMPARATIVE ANALYSIS --- p.65 / Chapter 4.1 --- PARTICIPANTS FLOW --- p.67 / Chapter 4.2 --- DEMOGRAPHICS --- p.69 / Chapter 4.3 --- BASELINE CHARACTERISTICS COMPARISON --- p.70 / Chapter 4.3.1 --- IPSS --- p.72 / Chapter 4.3.2 --- QMAX AND PRV --- p.73 / Chapter 4.3.3 --- QUALITY OF LIFE --- p.73 / Chapter 4.3.4 --- SERUM PSA LEVEL --- p.74 / Chapter 4.4 --- EFFICACY OUTCOMES --- p.74 / Chapter 4.4.1 --- QMAX AND PVR --- p.74 / Chapter 4.4.1.1 --- QMAX --- p.74 / Chapter 4.4.1.2 --- PVR --- p.75 / Chapter 4.4.2 --- IPSS --- p.79 / Chapter 4.4.2.1 --- TOTAL IPSS --- p.79 / Chapter 4.4.2.2 --- IPSS SUB SCORE 1_ INCOMPLETE EMPTYING --- p.79 / Chapter 4.4.2.3 --- IPSS SUB SCORE 2_ FREQUENCY --- p.80 / Chapter 4.4.2.4 --- IPSS SUB SCORE 3_INTERMITTENCY --- p.81 / Chapter 4.4.2.5 --- IPSS SUB SCORE 4_ URGENCY --- p.82 / Chapter 4.4.2.6 --- IPSS SUB SCORE 5_ WEAK STREAM --- p.82 / Chapter 4.4.2.7 --- IPSS SUB SCORE 6_ STRAINING --- p.83 / Chapter 4.4.2.8 --- IPSS SUB SCORE 7_ NOCTURIA --- p.84 / Chapter 4.4.3 --- QOL --- p.90 / Chapter 4.4.3.1 --- QOL IN IPSS Q8_QOL_URINATION --- p.90 / Chapter 4.4.3.2 --- QOL IN SF-36 --- p.91 / Chapter 4.4.3.2.1 --- PHYSICAL FUNCTIONING --- p.91 / Chapter 4.4.3.2.2 --- ROLE-PHYSICAL --- p.92 / Chapter 4.4.3.2.3 --- BODY PAIN --- p.92 / Chapter 4.4.3.2.4 --- GENERAL HEALTH --- p.93 / Chapter 4.4.3.2.5 --- VITALITY --- p.94 / Chapter 4.4.3.2.6 --- SOCIAL FUNCTIONING --- p.95 / Chapter 4.4.3.2.7 --- ROLE-EMOTIONAL --- p.95 / Chapter 4.4.3.2.8 --- MENTAL HEALTH --- p.96 / Chapter 4.4.4 --- SERUM PSA LEVEL --- p.103 / Chapter 4.4.5 --- SUBGROUP ANALYSIS --- p.106 / Chapter 4.4.6 --- SELF-PREFERENCE EFFECT ANALYSIS --- p.107 / Chapter 4.4.7 --- DIARY ANALYSIS --- p.109 / Chapter 4.5 --- ADVERSE EVENTS --- p.113 / Chapter 4.5.1 --- SELF-REPORTED AE/SAE --- p.113 / Chapter 4.5.2 --- SEXUAL HORMONE LEVEL --- p.114 / Chapter 4.5.3 --- SEXUAL RELATED QUALITY OF LIFE --- p.114 / Chapter 4.5.3.1 --- SEXUAL LIFE UNSATISFACTORY --- p.114 / Chapter 4.5.3.2 --- LIBIDO DECREASE --- p.115 / Chapter 5.1 --- PRINCIPAL FINDINGS --- p.116 / Chapter 5.1.1 --- EFFICACY --- p.116 / Chapter 5.1.2 --- SAFETY --- p.117 / Chapter 5.2 --- STRENGH AND LIMITATIOINS --- p.117 / Chapter 5.2.1 --- STRENGTH --- p.117 / Chapter 5.2.1.1 --- BLINDING IS EFFECTIVE --- p.117 / Chapter 5.2.1.2 --- COMPLIANCE IS GOOD --- p.118 / Chapter 5.2.1.3 --- LONG TREATMENT PERIOD --- p.119 / Chapter 5.2.1.4 --- STUDY OUTCOMES INCLUDE BOTH OBJECTIVE OUTCOMES AND SUBJECTIVE OUTCOMES --- p.121 / Chapter 5.2.2 --- LIMITATIONS --- p.121 / Chapter 5.2.2.1 --- INSUFFICIENT SAMPLE SIZE --- p.122 / Chapter 5.2.2.2 --- POSSIBLY LOW DOSE --- p.122 / Chapter 5.2.2.3 --- LACK OF BASELINE DATA ON QUALITY OF SEXUAL LIFE --- p.123 / Chapter 5.2.2.4 --- "LACK OF DATA ON LIFESTY FACTORES INCLUDING DIETARY HABIT, PHYSICAL ACTIVITY, SMOKING STATUS AND ACOHOL CONSUMPTION" --- p.123 / Chapter 5.3 --- INTERPRETATIONS OF THE RESUTLS --- p.124 / Chapter 5.3.1 --- TWO POSIVE RESULTS IN EMPTYING FUNCTION AND QUALITY OF LIFE --- p.124 / Chapter 5.3.2 --- ONE NEGATIVE RESULT IN PEAK URINARY FLOW RATE --- p.127 / Chapter 5.3.3 --- QUALITY OF SEXUAL LIFE --- p.129 / Chapter 6.1 --- CONCLUSIONS --- p.130 / Chapter 6.2 --- IMPLICATIONS --- p.131

Yuen, Ka Leung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 95-104). / Abstracts in English and Chinese. / Chapter 1 --- Introduction / Chapter 1.1 --- CANCER / Chapter 1.1.1 --- OVERVIEW OF CANCER --- p.1 / Chapter 1.1.2 --- DEVELOPMENT OF CANCER --- p.1 / Chapter 1.1.3 --- CHARACTERISTICS OF CANCER CELLS --- p.3 / Chapter 1.1.4 --- CATEGORIZATION OF CANCER --- p.6 / Chapter 1.1.5 --- RISK FACTORS IN CANCER DEVELOPMENT --- p.7 / Chapter 1.1.6 --- EPIDEMIOLOGY OF CANCER --- p.11 / Chapter 1.1.7 --- CANCER THERAPIES --- p.13 / Chapter 1.2 --- SOYBEANS AND ISOFLAVONES / Chapter 1.2.1 --- GENERAL INTRODUCTION OF ISOFLAVONES --- p.18 / Chapter 1.2.2 --- NATURAL FUNCTIONS OF ISOFLAVONES --- p.19 / Chapter 1.2.3 --- STRUCTURES OF ISOFLAVONES --- p.19 / Chapter 1.2.4 --- BIOACTIVITIES OF SOY ISOFLAVONES --- p.20 / Chapter 1.2.5 --- PRODUCTION OF SOY ISOFLAVONES CAN BE AFFECTED BY MANY FACTORS --- p.21 / Chapter 1.3 --- THE AIM AND OBJECTIVES OF THE PROJECT / Chapter 1.3.1 --- AIM OF THE PROJECT --- p.22 / Chapter 1.3.2 --- OBJECTIVES OF THE PROJECT --- p.23 / Chapter 2 --- Materials / Chapter 2.1 --- 19 DROUGHT TOLERANT SOYBEAN LINES --- p.24 / Chapter 2.2 --- 5 HUMAN CANCER CELL LINES --- p.25 / Chapter 2.3 --- CHEMICALS --- p.25 / Chapter 2.4 --- REAGENTS --- p.26 / Chapter 2.5 --- SOLUTIONS --- p.26 / Chapter 2.6 --- MAJOR EQUIPMENTS AND MATERIALS --- p.28 / Chapter 3 --- Methodology / Chapter 3.1 --- PREPARATION OF SOYBEAN EXTRACTS --- p.29 / Chapter 3.2 --- HIGH PERFORMANCE LIQUID CHROMATOGRAPHY(HPLC) ANALYSIS OF SOYBEAN EXTRACTS / Chapter 3.2.1 --- PREPARATION OF SOYBEAN EXTRACTS FOR HPLC ANALYS --- p.30 / Chapter 3.2.2 --- HPLC ANALYSIS --- p.30 / Chapter 3.3 --- PREPARATION OF 5 HUMAN CANCER CELL LINES FOR ANTI-PROLIFERATION ASSAY / Chapter 3.3.1 --- THAWING OF THE C Y R O P R E S E R V E D CELL LINES --- p.31 / Chapter 3.3.2 --- MAINTAINING OF CELL LINES --- p.32 / Chapter 3.3.3 --- ANTI-PROLIFERATION TEST WITH MTT ASSAY --- p.33 / Chapter 3.4 --- STATISTICS --- p.35 / Chapter 4 --- Results / Chapter 4.1 --- PREPARARTION OF SOYBEAN EXTRACTS --- p.36 / Chapter 4.2 --- HPLC ANALYSIS OF 5 SELECTED ISOFLAVONES IN 19 SOYBEAN SAMPLES --- p.36 / Chapter 4.3 --- COMPARISON OF SUM OF 5 SELECTED ISOFLAVONES FROM THE PARENT SOYBEAN AND VARIETIES HARVESTED FROM IRRIGATED LAND --- p.43 / Chapter 4.4 --- COMPARISON OF SUM OF 5 SELECTED ISOFLAVONES FROM THE SOYBEAN VARIETIES HARVESTED FROM DROUGHT LAND --- p.45 / Chapter 4.5 --- COMPARISON OF SELECTED ISOFLAVONES FROM THE PARENT SOYBEAN AND VARIETIES HARVESTED FROM IRRIGATED LAND --- p.47 / Chapter 4.6 --- COMPARISON OF SELECTED ISOFLAVONES FROM THE PARENT SOYBEAN AND VARIETIES HARVESTED FROM DAROUGHT LAND --- p.54 / Chapter 4.7 --- COMPARISON OF SUM OF SELECTED ISOFLAVONES AMONG THE SOYBEANS HARVESTED FROM IRRIGATED LAND AND DROUGHT LAND --- p.59 / Chapter 4.8 --- DETERMINATION OF ANTI-PROLIFERATION ABILITIES OF SOYBEAN SAMPLES --- p.19 / Chapter 4.8.1 --- ANTI-PROLIFERATION TEST OF ETHANOL AND 2-PHEN YLCHROMONE --- p.61 / Chapter 4.8.2 --- ANTI-PROLIFERATION ACTIVITIES OF 19 SOYBEAN SAMPLES ON 5 HUMAN CANCER CELL LINES --- p.61 / Chapter 4.9 --- COMPARISON OF ANTI-PROLIFERATION POTENCIES OF19 SOYBEAN SAMPLES WITH SUM OF SELECTED ISOFLAVONES --- p.70 / Chapter 4.10 --- COMPARISON OF ANTI-PROLIFERATION POTENCIES OF19 SOYBEAN SAMPLES --- p.72 / Chapter 4.11 --- ANTI-PROLIFERATION EFFECT OF INDIVIDUAL ISOFLAVONES ON FIVE CANCER CELL LINES --- p.74 / Chapter 5 --- Discussion / Chapter 5.1 --- EXTRACTION OF 19 SOYBEAN LINES --- p.77 / Chapter 5.2 --- DETERMINATION OF QUANTITIES OF SELECTED ISOFLAVONES IN 19 SOYBEAN SAMPLES BY HPLC ANALYSIS --- p.77 / Chapter 5.3 --- COMPARISON OF SELECTED ISOFLAVONES AMONG 19 SOYBEAN SAMPLES / Chapter 5.3.1 --- COMPARISON OF SUM OF SELECTED ISOFLAVONES BETWEEN PARENT AND SOYBEANS HARVESTED FROM IRRIGATED LAND --- p.80 / Chapter 5.3.2 --- COMPARISON OF SUM OF SELECTED ISOFLAVONES BETWEEN SOYBEANS HARVESTED FROM DROUGHT LAND --- p.81 / Chapter 5.3.3 --- COMPARISON OF SELECTED ISOFLAVONES BETWEEN SOYBEANS HARVESTED FROM IRRIGATED LAND --- p.81 / Chapter 5.3.4 --- COMPARISON OF SELECTED ISOFLAVONES BETWEEN SOYBEANS HARVESTED FROM DROUGHT LAND --- p.82 / Chapter 5.3.5 --- COMPARISON OF SUM OF SELECTED ISOFLAVONES BETWEEN SOYBEANS HARVESTED FROM IRRIGATED LAND AND DROUGHT LAND --- p.83 / Chapter 5.4 --- COMPARISON OF ANTI-PROLIFERATION ACTIVITIES OF 19 SOYBEAN SAMPLES / Chapter 5.4.1 --- COMPARISON OF ANTI-PROLIFERATION ACTIVITIES OF19 SOYBEAN SAMPLES AMONG 5 CANCER CELL LINES --- p.84 / Chapter 5.4.2 --- COMPARISON OF ANTI-PROLIFERATION POTENCIES OF19 SOYBEAN SAMPLES --- p.85 / Chapter 5.4.3 --- COMPARISON OF ANTI-PROLIFERATION ACTIVITIES OF19 SOYBEAN SAMPLES AND CORRESPONDING SUM OF SELECTED ISOFLAVONES --- p.86 / Chapter 5.4.4 --- COMPARISON OF IC50S FROM SOYBEANS HARVESTED FROM IRRIGATED LAND AND DROUGHT LAND --- p.87 / Chapter 5.4.5 --- CORRELATION OF ISOFLAVONES AND ANTI-PROLIFERATION POTENCIES --- p.88 / Chapter 6 --- Conclusion --- p.90 / Chapter 7 --- References --- p.91 / Chapter 8 --- Appendix --- p.S1

Yuen Yee Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 108-125). / Abstracts in English and Chinese. / TITLE PAGE --- p.1 / ACKNOWLEGDEMENTS --- p.2 / ABSTRACT --- p.3 / 摘要 --- p.5 / table of contents --- p.7 / list of figures and tables --- p.13 / CHAPTER 1 GENERAL INTRODUCTION --- p.16 / Chapter 1.1 --- role of PHYTOESTROGENS in soy and red WINE the PREVENTION OF CARDIOVASCULAR DISEASES (CVD) --- p.17 / Chapter 1.1.1 --- INTRoduction and Classification of Phytoestrogens --- p.17 / Chapter 1.1.2 --- estrogenic1ty of phytoestrogens and theIr abundancesin Plasma --- p.18 / Chapter 1.1.3 --- phytoestrogens as one of the active components In cvd Protection --- p.21 / Chapter 1.1.4 --- effects of Phytoestrogens on LDL Receptor and Apolipoprotein A-1 --- p.22 / Chapter 1.2 --- role of estrogen receptors (ers) in gene regulation --- p.24 / Chapter 1.2.1 --- "structure, Classification and tissue distribution of ERS" --- p.24 / Chapter 1.2.2 --- ligands for ERS --- p.25 / Chapter 1.2.3 --- mechaniSMS OF LIgands-ERS complex in GENE regulation --- p.27 / Chapter 1.2.4 --- ligand-independent ER activation --- p.28 / Chapter 1.3 --- aims and scopes of investigation --- p.29 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.30 / Chapter 2.1 --- chemicals and materials --- p.30 / Chapter 2.1.1 --- Chemicals --- p.30 / Chapter 2.1.2 --- Plasmids --- p.30 / Chapter 2.2 --- mammalian cell culture maintainence --- p.30 / Chapter 2.2.1 --- Maintenance of Cells --- p.31 / Chapter 2.2.2 --- Preparation of Cell Stock --- p.31 / Chapter 2.2.3 --- Cell Recovery from Liquid Nitrogen Stock --- p.31 / Chapter 2.3 --- manipulation of dna --- p.31 / Chapter 2.3.1 --- isolation of HEPG2 cells genonmic DNA --- p.31 / Chapter 2.3.2 --- separation and purification of dna from agarose gel --- p.31 / Chapter 2.3.3 --- Restriction digestionof DNA --- p.32 / Chapter 2.3.4 --- Ligation of DNA Fragments --- p.32 / Chapter 2.3.5 --- Transformation of --- p.32 / Chapter 2.3.6 --- Small Scale Plasmids Purification from DH5a --- p.32 / Chapter 2.4 --- construction of expression and reporter plasmids --- p.33 / Chapter 2.4.1 --- Construction of Estrogen Receptorα (Erα) Expression Vectors --- p.33 / Chapter 2.4.2 --- construction of reporter vectors of LDLR promoter and the Respective Mutants --- p.33 / Chapter 2.4.3 --- Construction of Reporter Vectors of APOAI Promoter and the Respective Mutants --- p.33 / Chapter 2.5 --- determination of promoter transcrtiption activities --- p.34 / Chapter 2.5.1 --- Transient Transfection of Cell with ERa Expression Vector and Promoter Reporter using Lipofectamine PLUS Reagent --- p.34 / Chapter 2.5.2 --- Dual Luciferase Assay --- p.34 / Chapter 2.6 --- semi-quantitative and quantitative rt-pcr assay --- p.34 / Chapter 2.6.1 --- Transient transfection of Cell with ERa Expression Vector Using Lipofectamine PLUS Reagent --- p.34 / Chapter 2.6.2 --- "Isolation of RNA using TRIzol® Reagent (Life Technology, USA)" --- p.35 / Chapter 2.6.3 --- Quantitation of RNA --- p.35 / Chapter 2.6.4 --- First Strand cDNA Synthesis --- p.35 / Chapter 2.6.5 --- Sem卜Quantitative PCR Reactions --- p.35 / Chapter 2.6.6 --- Quantitative PCR Reactions --- p.36 / Chapter 2.7 --- western blotting analysis --- p.36 / Chapter 2.8 --- statistical methods --- p.36 / Chapter CHAPTER 3 --- REGULATION BY PHYSIOLOGICAL LEVEL OF 17B-ESTRADIOL ON APOLIPOPROTEIN A-I AND LOW-DENSITY- LIPOPROTEIN RECEPTOR IN HEPG2 CELLS --- p.37 / Chapter 3.1 --- introduction --- p.37 / Chapter 3.2 --- results --- p.39 / Chapter 3.2.1 --- Determination of transient transfection functionality of estrogen receptors in hepg2 cells --- p.39 / Chapter 3.2.2 --- Effect of 17β-Estradiolon LDLR promoter transcription activity --- p.39 / Chapter 3.2.3 --- Effect of 17β-Estradiol on apoai promoter transcription activity --- p.40 / Chapter 3.2 --- discussion --- p.47 / Chapter CHAPTER 4 --- SOY ISOFLAVONES AND RESVERATROL DISPLAY DIFFERENT MECHANISM IN THE UP-REGULATION OF LOVV-DENSITY-LIPOPROTEIN RECEPTOR IN HEPG2 CELLS --- p.49 / Chapter 4.1 --- introduction --- p.49 / Chapter 4.2 --- results --- p.54 / Chapter 4.2.1 --- Association of ERα and isoflavones or resveratrol on LDLR promoter transcription activity --- p.54 / Chapter 4.2.2 --- Association of ERβ and isoflavones or resveratrol on LDLR promoter transcription activity --- p.54 / Chapter 4.2.3 --- "Role of MAP Kinase, PKA and PKC in isoflavones and resveratrol induced LDLR promoter transcription" --- p.55 / Chapter 4.2.4 --- Identification of promoter regions responsible for induction of LDLR transcription by isoflavones in the presence OF ERα --- p.55 / Chapter 4.2.5 --- Identification of promoter regions responsible for induction of LDLR TRANSCRIPTION BY resveratrol IN THE ABSENCE OF ERα --- p.56 / Chapter 4.3 --- DISCUSSION --- p.75 / Chapter CHAPTER 5 --- SOY ISOFLAVONES AND RESVERATROL UP-REGULATE APOLIPOPROTEIN A-I SIMILAR TO 17B-ESTRADIOL IN HEPG2 CELLS --- p.80 / Chapter 5.1 --- INTRODUCTION --- p.80 / Chapter 5.2 --- RESULTS --- p.84 / Chapter 5.2.1 --- Association of ERα phytoestrogens on APCAI gene expression --- p.84 / Chapter 5.2.2 --- Association of ERβ and isoflavones or resveratrol on APOAI promoter transcription activity --- p.85 / Chapter 5.2.3 --- "Role of MAP Kinase, PKA and PKC in isoflavones and resveratrol in APOAI promoter transcription in the presence of ERα" --- p.85 / Chapter 5.2.4 --- Identification of promoter regions responsible for induction of APOAI transcription by isoflavones and resveratrol in the presence of ERα --- p.85 / Chapter 5.3 --- DISCUSSION --- p.100 / Chapter CHAPTER 6 --- GENERAL DISCUSSION --- p.103 / Chapter CHAPTER 7 --- SUMMARY --- p.106 / BIBLIOGRAPHY --- p.108 / APPENDIX 1 ABBREVIATIONS --- p.126 / APPENDIX 2 MATERIALS AND METHODS --- p.129 / APPENDIX 3 PRIMER LISTS --- p.145 / APPENDIX 4 REAGENTS AND BUFFERS --- p.147