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An expression and functional study of an orphan nuclear receptor, estrogen receptor-related receptor (ERR), in the human prostate and prostate cancer.

Cheung Chun Pan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 192-227). / Abstracts in English and Chinese. / Acknowledgements --- p.I / Abstract (English) --- p.II / Abstract (Chinese) --- p.VI / Contents --- p.VIII / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Nuclear hormone receptor: a general review --- p.1 / Chapter 1.1.1 --- Classification of nuclear hormone receptors --- p.1 / Chapter 1.1.2 --- Mechanism of action --- p.2 / Chapter 1.1.3 --- Domains structure and functions --- p.3 / Chapter 1.1.4 --- Orphan nuclear receptors --- p.4 / Chapter 1.2 --- Prostate gland - a male accessory reproductive organ --- p.6 / Chapter 1.2.1 --- "Anatomy, histology and physiology of the prostate gland" --- p.6 / Chapter 1.2.2 --- Endocrinology of the prostate gland --- p.8 / Chapter 1.2.3 --- Pathogenesis of the prostate gland --- p.8 / Chapter 1.3 --- The role of estrogen receptors in the prostate gland and prostate cancer --- p.9 / Chapter 1.3.1 --- Estrogens in male --- p.10 / Chapter 1.3.2 --- Effects of estrogens in the prostate gland --- p.11 / Chapter 1.3.3 --- Estrogen receptors - two isoforms --- p.13 / Chapter 1.3.4 --- Expression of ERs in the prostate gland --- p.14 / Chapter 1.3.5 --- Estrogen-modulated transgenic mice 226}0ؤ functional studies of ERs --- p.16 / Chapter 1.4 --- Estrogen receptor-related receptors: orphan receptors --- p.18 / Chapter 1.4.1 --- Estrogen receptor-related receptors: Three isoforms --- p.18 / Chapter 1.4.2 --- Expression of ERRs in different tissues --- p.20 / Chapter 1.4.3 --- Promoter binding and genes regulated by of ERRs --- p.22 / Chapter 1.4.4 --- Coregulators of ERRs --- p.24 / Chapter 1.4.5 --- Ligand of ERRs --- p.25 / Chapter 1.4.6 --- Functional roles of ERRs --- p.29 / Chapter 1.4.7 --- Cross talk between ERRs and ERs --- p.31 / Table 11 --- p.33 / Figure 1.1 - 15 --- p.35 / Chapter Chapter 2 --- Aims of the Study --- p.40 / Chapter Chapter 3 --- Methods and Materials / Chapter 3.1 --- "Expression patterns of ERRs and steroid hormone receptors in the human prostate cell lines, tumor xenografts and prostatic tissues" --- p.41 / Chapter 3.1.1 --- Human prostatic tissues --- p.41 / Chapter 3.1.2 --- Cell cultures --- p.41 / Chapter 3.1.3 --- Human prostate cancer xenografts --- p.42 / Chapter 3.1.4 --- Full length clones of ERR isoforms --- p.42 / Chapter 3.1.5 --- Reverse transcription-polymerase chain reactions (RT-PCR) --- p.43 / Chapter 3.1.6 --- Semi-quantitative RT-PCR analysis --- p.45 / Chapter 3.1.7 --- Southern blot analysis --- p.46 / Chapter 3.1.8 --- Generation and characterization of polyclonal antibodies --- p.49 / Chapter 3.1.9 --- Western blot analysis --- p.55 / Chapter 3.1.10 --- Immunohistochemistry --- p.56 / Chapter 3.2 --- Relationship of ERR and ER expressions in the prostatic cells --- p.57 / Chapter 3.2.1 --- "Expression vectors of ERRa, ERRγ and ERa" --- p.57 / Chapter 3.2.2 --- "Transient transfection of ERRa, ERRγ and ERa expression vectors in PC-3 cells" --- p.58 / Chapter 3.2.3 --- Semi-quantitative RT-PCR analysis --- p.59 / Chapter 3.3 --- Intracellular trafficking and transcriptional activity of GFP-tagged ERRs in the prostatic cells --- p.59 / Chapter 3.3.1 --- Construction of GFP-tagged ERR fusion plasmids --- p.59 / Chapter 3.3.2 --- Examination of transcriptional activity of GFP-tagged ERRs by luciferase assay --- p.61 / Chapter 3.3.3 --- Subcellular localization of GFP-tagged ERRs in the living prostatic cells --- p.63 / Chapter 3.3.4 --- Immunofluorescent staining GFP-tagged ERRs --- p.63 / Chapter 3.4 --- The Role of ERRs in the growth of the prostatic cells --- p.64 / Chapter 3.4.1 --- Evaluation the transfection efficiencies of PC-3 and PNT2 cells --- p.64 / Chapter 3.4.2 --- Cells proliferation assays in ERRs transient transfected prostatic cells --- p.66 / Chapter 3.4.3 --- Flow cytometry of ERRs transient transfected PC-3 cells --- p.66 / Chapter 3.4.4 --- RT-PCR of cell cycle-related genes in ERRs transient transfected PC-3 cells --- p.67 / Chapter 3.4.5 --- Generation of PNT2 and DU145 cells stably transfected with ERRy --- p.68 / Chapter 3.4.6 --- Cell proliferation assay of ERRy stable-transfected PNT2 and DU 145cells --- p.72 / Chapter 3. 4.7 --- Anchorage independent growth assay of ERRy stable-transfected PNT2 and DU 145 cells --- p.72 / Chapter 3.4.8 --- Flow cytometry of ERRγ stable-transfected PNT2 and DU145 cells --- p.74 / Chapter 3.4.9 --- RT-PCR of cell cycle-related genes in ERRγ stable-transfected PNT2 and DU 145 cells --- p.74 / Chapter 3.4.10 --- Western blot analysis of p21 in ERRγ stable-transfected PNT2 cells --- p.75 / Chapter 3.5 --- Statistical analysis --- p.75 / Table 3.1 - 3.2,Figure 3.1 - 35 --- p.76 / Chapter Chapter 4 --- Results / Chapter 4.1 --- "Expression patterns of ERRs and steroid hormone receptors in the human prostate cell lines, tumor xenografts and prostatic tissues" --- p.93 / Chapter 4.1.1 --- "mRNA expression patterns of ERR isoforms in the prostatic cell lines, prostate cancer xenografts and human prostatic tissues" --- p.93 / Chapter 4.1.2 --- mRNA expression patterns of steroid hormone receptors and prostatic differentiation markers in the prostatic cell lines and xenografts --- p.95 / Chapter 4.1.3 --- Characterization of antisera against human ERRs by ERR recombinant proteins --- p.97 / Chapter 4.1.4 --- Protein expression of ERR isoforms in the human prostatic cell lines --- p.98 / Chapter 4.1.5 --- "Immunolocalization of ERR isoforms in the normal, dysplastic and neoplastic prostates" --- p.98 / Chapter 4.2 --- Interrelationship of ERR and ER expression in PC-3 prostate cancer cells --- p.100 / Chapter 4.2.1 --- "Expressions of ERRγ, ERa and ERβ in the ERRa transient transfected PC-3 cells" --- p.100 / Chapter 4.2.2 --- Expression of ERRa in the ERRγ and ERa transient transfected PC-3 cells --- p.101 / Chapter 4.3 --- Intracellular trafficking and transcriptional activities of ERRs in the prostatic cells with fused green fluorescence protein 一 ERRs --- p.102 / Chapter 4.3.1 --- Trans activation of ERE response element 226}0ؤ driven reporter by ERR isoforms in the PC-3 cells in the presence or absence of serum --- p.102 / Chapter 4.3.2 --- Trans activation of SF-1 response element driven reporter by ERR isoforms in the PC-3 cells in the presence or absence of serum --- p.104 / Chapter 4.3.3 --- Subcellular localizations of three ERR isoforms in the PC-3 cells in the presence or absence of serum --- p.105 / Chapter 4.4 --- The role of ERRs in the growth of prostatic cells --- p.106 / Chapter 4.4.1 --- "The growth of ERRs transient transfected PC-3, PNT2 prostatic cells" --- p.107 / Chapter 4.4.2 --- Cell cycle analysis of ERRs transient transfected PC-3 cells --- p.108 / Chapter 4.4.3 --- Expression of cyclin-dependent kinase (CDK) inhibitors and p53 in the ERRs transient transfected PC-3 cells --- p.108 / Chapter 4.4.4 --- Establishment of ERRγ stable-transfected PNT2 and DU145 cells --- p.109 / Chapter 4.4.5 --- Transcriptional activation of ERE response element in ERRγ stable-transfected PNT2 cells --- p.111 / Chapter 4.4.6 --- Effect of over-expression of ERRγ on the growth of PNT2 and DU145 stable-transfected cells --- p.112 / Chapter 4.4.7 --- Efficiencies of colony formation of ERRγ stable-transfected PNT2 and DU145 cells --- p.113 / Chapter 4.4.8 --- Cell cycle analysis of ERRγ stable-transfected PNT2and DU 145 cells --- p.114 / Chapter 4.4.9 --- Expression of cell cycle-related genes in the ERRy stable-transfected PNT2 and DU 145 cells --- p.116 / Figure 4.1 - 4.38,Table 4.1 - 43 --- p.119 / Chapter Chapter 5 --- Discussion / Chapter 5.1 --- "Expression study in human prostatic cells, tumor xenografts" --- p.159 / Chapter 5.1.1 --- "Differential expression patterns of ERRs in prostatic cells, cancer xenografts and tissues" --- p.160 / Chapter 5.1.2 --- Co-localization of ERRs and ERβ in the human prostate --- p.166 / Chapter 5.1.3 --- Differential expression patterns of steroid hormone receptors and prostatic specific markers in prostatic cells and xenografts --- p.168 / Chapter 5.2 --- ERRα acts as a expression repressor of ERRγ and ERα in PC-3 cells --- p.173 / Chapter 5.3 --- ERRs are nuclear localized and constitutively active in PC-3 cells --- p.176 / Chapter 5.4 --- ERRs acts as the negative growth regulators in the prostatic cells --- p.179 / Chapter 5.4.1 --- Cell cycle control of mammalian cells --- p.180 / Chapter 5.4.2 --- The roles of AR and ERs in the cell cycle regulation --- p.181 / Chapter 5.4.3 --- Inhibition of cell proliferation in ERRs transient transfected PC-3 cells and ERRγ stable-transfected PNT2 and DU145 cells --- p.184 / Chapter 5.4.4 --- Inhibition of anchorage independent growth in ERRγ stable-transfected PNT2 and DU 145 cells --- p.188 / Chapter Chapter 6 --- Conclusion --- p.191 / Chapter Chapter 7 --- References --- p.192 / Chapter Chapter 8 --- Publications --- p.227

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324633
Date January 2004
ContributorsCheung, Chun Pan., Chinese University of Hong Kong Graduate School. Division of Anatomy.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xv, 227 leaves, [38] leaves of plates : ill. (some col.) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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