Chan, Ming Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 92-106). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.ii / 摘要 --- p.iv / LIST OF ABBREVIATIONS --- p.vi / TABLE OF CONTENTS --- p.viii / Chapter CHAPTER 1 --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Aromatase --- p.1 / Chapter 1.2 --- Tissue Specific Promoter for Aromatase Expression --- p.4 / Chapter 1.3 --- Signaling Pathway --- p.7 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.9 / Chapter 2.1 --- Chemicals And Materials --- p.9 / Chapter 2.2 --- Mammalian Cell Culture --- p.9 / Chapter 2.2.1 --- Maintenance of Cells --- p.10 / Chapter 2.2.2 --- Preparation of Cells Stock --- p.10 / Chapter 2.2.3 --- Cell Recovery from Liquid Nitrogen Stock --- p.11 / Chapter 2.3 --- Tritiated Water Release Assay --- p.11 / Chapter 2.3.1 --- Aromatase Activity in Intact Cell --- p.11 / Chapter 2.3.2 --- Aromatase Assay on Recombinant Supersomes --- p.12 / Chapter 2.4 --- RNA Isolation and cDNA Synthesis --- p.13 / Chapter 2.5 --- Semi-Quantitative PCR Reaction --- p.13 / Chapter 2.6 --- Quantitative Real Time PCR Using Taqman Probe --- p.15 / Chapter 2.7 --- Western Blotting --- p.17 / Chapter 2.8 --- Measurement of Promoter Activity --- p.18 / Chapter 2.8.1 --- Plasmid Preparation --- p.18 / Chapter 2.8.2 --- Transient Transfection and Dual Luciferase Assay --- p.18 / Chapter 2.9 --- Statistical Methods --- p.19 / Chapter CHAPTER 3 --- Genistein up-regulate aromatase in Estrogen receptor alpha-transfected HepG2 cells --- p.21 / Chapter 3.1 --- Introduction --- p.21 / Chapter 3.1.1 --- Cardiovascular Disease (CVD) --- p.21 / Chapter 3.1.2 --- Phytoestrogen --- p.21 / Chapter 3.1.3 --- Estrogen Receptor --- p.24 / Chapter 3.1.4 --- Protective Mechanism Against CVD Protection --- p.25 / Chapter 3.1.5 --- Effects of genistein on LDL Receptor and Apolipoprotein A-I --- p.26 / Chapter 3.1.6 --- Effects of estradiol on LDL Receptor and Apolipoprotein A-I　 --- p.26 / Chapter 3.1.7 --- Aim of study and hypothesis --- p.27 / Chapter 3.2 --- Result --- p.29 / Chapter 3.2.1 --- ERa increased Aromatase Activity in HepG2 cells --- p.29 / Chapter 3.2.2 --- Genistein increased Aromatase Activity in HepG2 cells --- p.29 / Chapter 3.2.3 --- Differential Effect of MAP kinase Inhibitors --- p.35 / Chapter 3.2.4 --- "Role of MAP Kinase, PKA and PKC in Genistein Induced Aromatase Activity in ERa-transfected HepG2 cells" --- p.35 / Chapter 3.2.5 --- Genistein Increased Aromatase Protein Expression in ERa-transfected HepG2 cells --- p.38 / Chapter 3.2.6 --- Genistein Induced Aromatase mRNA Expression Attributed to Induction of Exon ̐ơ.1 Expression --- p.40 / Chapter 3.2.7 --- Genistein Induced Promoter 1.1 Transcriptional Activity in ERa- transfected HepG2 cells --- p.44 / Chapter 3.2.8 --- Genistein Increased ERE and AP-1 Reporter Activity Through Interaction with ERa --- p.47 / Chapter 3.3 --- Discussion --- p.51 / Chapter CHAPTER 4 --- "Effect of 2,3,7,8-tetrachlorodibenzo- para-TCDD (TCDD) on aromatase in MCF-7 cells" --- p.54 / Chapter 4.1 --- Introduction --- p.54 / Chapter 4.1.1 --- Breast Cancer --- p.54 / Chapter 4.1.2 --- TCDD --- p.54 / Chapter 4.1.3 --- CYP Enzymes --- p.55 / Chapter 4.1.4 --- TCDD and Breast Cancer --- p.56 / Chapter 4.1.5 --- Aim of Study --- p.56 / Chapter 4.2 --- Result --- p.57 / Chapter 4.2.1 --- Effect of TCDD on Aromatase Activity in Different Cell Lines --- p.57 / Chapter 4.2.2 --- TCDD Increased Aromatase Activity in MCF-7 Cells --- p.62 / Chapter 4.2.3 --- Effect of TCDD on Human CYP 19 Recombinant Supersomes® and MCF-7aro Cells --- p.66 / Chapter 4.2.4 --- TCDD Increased Aromatase Protein Expression in MCF-7 Cells --- p.66 / Chapter 4.2.5 --- Effect of TCDD in Aromatase mRNA Expression in MCF-7 Cells --- p.70 / Chapter 4.2.6 --- Effect of TCDD in CYP 19 Promoter and AP-1 Promoter Activity in MCF-7 Cells --- p.70 / Chapter 4.2.7 --- Effect of TCDD in CYP 19 mRNA Half-life --- p.75 / Chapter 4.2.8 --- "Role of MAP Kinase, PKA and PKC in Genistein Induced Aromatase Activity in MCF-7 Cells" --- p.78 / Chapter 4.2.9 --- TCDD induced ERK1/2 Activation --- p.78 / Chapter 4.2.10 --- Induction of aromatase activity in MCF-7erk cells --- p.78 / Chapter 4.3 --- Discussion --- p.87 / Chapter CHAPTER 5 --- Summary --- p.90 / BIBLIOGRAPHY --- p.92

Graduation date: 1998

Thesis (M.S.P.)--University of Toledo, 2004. / Typescript. "A thesis [submitted] as partial fulfillment of the requirements of the Master of Science degree in Pharmaceutical Sciences." Includes bibliographical references (leaves 34-37).

Thesis (Ph.D.)--Washington State University, May 2006. / Includes bibliographical references (p. 141-160).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1984. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

No description available.

No description available.

Graduation date: 1999

The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure on hepatic pyruvate carboxylase (PC) gene expression were investigated in C57BL/6J Ah[superscipt b/b male mice. A dose-dependent reduction of PC levels and activity occurred in animals given a single intraperitoneal dose of TCDD in a corn oil carrier. The dose ranged from 1 to 75 ug/kg body weight and the analysis performed 8 days postinjection. At the maximum TCDD level investigated, a 10-fold reduction in PC activity occurred. At doses beyond those required to initiate a reduction in PC, a lactate dehydrogenase isozyme patterns shift is observed. This is accompanied by increases in blood lactic acid levels. Northern blot analysis on RNA extracts from hepatic tissues indicated that at 8 days post TCDD treatment, a dose-dependent reduction of hepatic mRNA levels occurs. The aryl hydrocarbon receptor (AhR) is believed to mediate all responses to TCDD. Liganded AhR and the aryl hydrocarbon receptor nuclear translocator (ARNT) protein form a heterodimeric transcription factor which interacts with dioxin response elements (DREs). These are found in enhancer/promoter regions of many genes that respond transcriptionally to TCDD exposure. Cloning and sequencing a region approximately 1.4 kb upstream of the PC translational start site revealed an untranslated leader sequence of 124 nucleotides starting with adenosine. Primary structural analysis of the upstream region revealed an 1nr element in place of a TATA element. Additional transcription factor elements were identified including: Spl, GCF, UBP-1, GRE, CREB, NF-1, HNF-4, TFII-I and E-boxes; DRE elements were notably lacking. A tandem series of 10 evenly spaced E-boxes, which bind ARNT homodimers, are each juxtaposed to a TFII-I element, possibly forming composite elements. Tertiary structure analysis revealed the positioning of nine composite elements displayed as a trio of phased elements. Transient transfections into Hepa lc1c7 cells, using a luciferase reporter gene under the transcriptional control of the PC upstream region, unlike the animal studies, produced an induction in activity in the presence of 10 nM TCDD. Co-transfections with an ARNT encoding plasmid reduced induction indicating overexpression of ARNT protein partially overrides the TCDD-induced increase in activity. These results in relationship to whole animal experiments are discussed. / Graduation date: 1997

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.