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Cai, Ganhui., Thesis (Ph.D.)--Chinese University of Hong Kong, 2013., Includes bibliographical references (leaves 160-192)., Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web., also in Chinese., http://library.cuhk.edu.hk/record=b5884413, Use 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/)

Tse, Ho Man., Thesis (M.Phil)--Chinese University of Hong Kong, 2011., Includes bibliographical references (leaves 166-173)., s in English and Chinese., Acknowledgements --- p.i, p.ii, 撮要 --- p.v, Table of Content --- p.vi, Chapter Chapter 1: --- Introduction --- p.1, Chapter 1.1 --- Basis of Prostate Cancer --- p.1, Chapter 1.1.1 --- Epidemiology and Risk Factors of Prostate Cancer --- p.1, Chapter 1.1.2 --- Pathology of Prostate Cancer --- p.2, Chapter 1.1.3 --- Treatment Approaches for Prostate Cancer --- p.4, Chapter 1.2 --- Sex Hormones and Prostate Cancer --- p.7, Chapter 1.2.1 --- Prostate Development --- p.7, Chapter 1.2.2 --- Involvement of Sex Hormones in Prostate Cancer --- p.8, Chapter 1.2.3 --- Molecular Mechanisms of Sex Hormones --- p.13, Chapter 1.2.4 --- Hormone Receptor Antagonists --- p.15, Chapter 1.3 --- Involvement of microRNAs in Cancer --- p.19, Chapter 1.3.1 --- Basis of microRNAs --- p.19, Chapter 1.3.2 --- Aberrant microRNA Expressions in Cancers --- p.23, Chapter 1.3.3 --- Current Understandings on Regulations of micro RN A Expressions --- p.26, Chapter 1.3.4 --- Regulation of miRNA Expressions by Hormones --- p.29, Chapter 1.4 --- "Effects of the Anti-estrogen ICI 182,780 on Prostate Cancer Cells" --- p.30, Chapter 1.4.1 --- "ICI 182,780 Inhibits Cell Growth ofDU145" --- p.30, Chapter 1.5 --- Objectives of Project --- p.32, Chapter Chapter 2: --- Materials --- p.34, Chapter 2.1 --- Bacteria Strain --- p.34, Chapter 2.2 --- Tissue Culture Media --- p.34, Chapter 2.3 --- Plasmids --- p.34, Chapter 2.4 --- Kits and Accessories --- p.35, Chapter 2.5 --- Reagents and Solutions --- p.36, Chapter 2.6 --- DNA Oligos --- p.38, Chapter 2.7 --- Equipments --- p.40, Chapter Chapter 3: --- Methods --- p.41, Chapter 3.1 --- Cell Culture Conditions --- p.41, Chapter 3.2 --- miRNA Expression Profiling of DU145 --- p.41, Chapter 3.2.1 --- RNA Isolation --- p.41, Chapter 3.2.2 --- miRNA Microarray Profiling ofDU145 : --- p.42, Chapter 3.2.2.1 --- Fluorescent Labeling of RNA and Microarray Hybridization --- p.42, Chapter 3.2.2.2 --- Scanning and Analysis of Signals --- p.46, Chapter 3.2.3 --- Confirming miR-765 Up-regulation by ICI with qRT-PCR --- p.46, Chapter 3.2.3.1 --- Assessing ERp Dependency in miR-765 Induction --- p.48, Chapter 3.2.4 --- Effects of ICI on ARHGEF11 Expression --- p.49, Chapter 3.2.4.1 --- Reverse Transcription of mRNA --- p.50, Chapter 3.2.4.2 --- Quantitative Real-Time PCR for Gene mRNA expression --- p.50, Chapter 3.3 --- Characterizing the Promoter Region of miR-765 --- p.52, Chapter 3.3.1 --- Cloning of miR-765 Promoter into pGL3-Basic Vector --- p.52, Chapter 3.3.1.1 --- PCR Amplification of miR-765 Putative Promoter Region --- p.52, Chapter 3.3.1.2 --- Ligation of the Amplified Regions in pGL3-Basic Vector --- p.55, Chapter 3.3.1.3 --- Transformation and Screening of pGL3-765 Plasmid --- p.57, Chapter 3.3.1.4 --- Preparation of pGL3-765 Plasmid DNA --- p.59, Chapter 3.3.2 --- Preparation of Truncated miR- 765 Promoter Clones --- p.60, Chapter 3.3.2.1 --- pGL3-765-Trunc#l --- p.61, Chapter 3.3.2.2 --- pGL3-765-Trunc#2 --- p.62, Chapter 3.3.2.3 --- pGL3-765-Trunc#3 --- p.62, Chapter 3.3.3 --- Assessing the miR- 765 Promoter Activities --- p.63, Chapter 3.3.3.1 --- Optimizing Transfection Conditions --- p.64, Chapter 3.3.3.2 --- Co-transfection of pGL3-765 and pRL-CMV into DU145 Cells.. --- p.64, Chapter 3.3.3.3 --- Measuring Luciferase Activities --- p.65, Chapter 3.3.4 --- Computational Prediction of Transcription Factor Binding Sites on miR-765 Promoter --- p.66, Chapter 3.4 --- Characterizing the Promoter Region of ARHGEF11.. --- p.67, Chapter 3.4.1 --- Cloning of ARHGEF11 Promoter into pGL3-Basic Vector (pGL3-ARH) --- p.67, Chapter 3.4.1.1 --- PCR Amplification of ARHGEF11 Putative Promoter Region --- p.67, Chapter 3.4.1.2 --- Ligation of the Amplified Regions in pGL3-Basic Vector --- p.68, Chapter 3.4.1.3 --- Preparation of Plasmid DNA --- p.69, Chapter 3.4.2 --- Preparation of Truncated ARHGEF11 Promoter Clones --- p.69, Chapter 3.4.2.1 --- pGL3-ARH-Trunc#l --- p.69, Chapter 3.4.2.2 --- pGL3-ARH-Trunc#2 --- p.70, Chapter 3.4.2.3 --- pGL3-ARH-Trunc#3 --- p.71, Chapter 3.4.3 --- Assessing ARHGEF11 Promoter Activities --- p.72, Chapter 3.5 --- Identifying Transcription Factor Binding Sites on ARHGEF11 Promoter with EMS A --- p.73, Chapter 3.5.1 --- Computational Prediction --- p.73, Chapter 3.5.2 --- Preparation of Biotinylated Probe for use in EMSA --- p.73, Chapter 3.5.3 --- Preparation of Specific Competitors --- p.74, Chapter 3.5.4 --- Preparation of DU145 Nuclear and Cytoplasmic Extracts --- p.75, Chapter 3.5.4.1 --- Preparation of Extracts --- p.75, Chapter 3.5.4.2 --- Measuring Protein Concentrations --- p.76, Chapter 3.5.5 --- EMSA Detection of Interaction between Protein and Probe --- p.76, Chapter 3.6 --- Assessing Biological Significances of miR-765 --- p.78, Chapter 3.6.1 --- Effects of ICI on DU145 Cells Growth --- p.79, Chapter 3.6.2 --- Effects of ICI on DU145 Migration Ability --- p.79, Chapter 3.6.2.1 --- Monolayer Wound Healing Assay --- p.79, Chapter 3.6.2.2 --- Transwell Migration Assay --- p.80, Chapter 3.6.3 --- Validating Functionality of Ectopic miR- 765 --- p.81, Chapter 3.6.3.1 --- miR-765 Recognition Sequence --- p.81, Chapter 3.6.3.2 --- Preparation of pMIR-765 vector --- p.82, Chapter 3.6.3.3 --- Ectopic Introduction of miR-765 into DU145 Cells --- p.84, Chapter 3.6.3.4 --- "Verifying Functionality, of Ectopic miR-765" --- p.84, Chapter 3.6.4 --- Effects of miR-765 on DU145 Growth --- p.86, Chapter 3.6.5 --- Effects of miR-765 on DU145 Migration Ability --- p.86, Chapter 3.7 --- Statistical Analysis --- p.87, Chapter Chapter 4: --- Results --- p.88, Chapter 4.1 --- "Identifying ICI 182,780-Regulated miRNA in DU145 Cells" --- p.88, Chapter 4.1.1 --- miRNA Expression Profiling of DU145 with Microarray --- p.88, Chapter 4.1.2 --- "Confirming Induction of miR-765 by ICI 182,780 with qRT-PCR" --- p.91, Chapter 4.1.3 --- "ARHGEF11, Host Gene of miR-765" --- p.95, Chapter 4.1.4 --- "Induction of ARHGEF 11 by ICI 182,780" --- p.96, Chapter 4.2 --- Characterization miR-765 Promoter Region --- p.98, Chapter 4.2.1 --- Cloning of miR- 765 Promoter Region into pGLS-Basic Vector --- p.98, Chapter 4.2.2 --- Promoter Activity of miR-765 Promoter --- p.100, Chapter 4.2.3 --- Deletion Mapping of miR- 765 Promoter Region --- p.102, Chapter 4.2.4 --- Promoter Activities and Inducibitiy of Truncated miR-765 Promoters --- p.103, Chapter 4.2.5 --- Computational Prediction of Transcription Factor Binding Sites on miR-765 Promoter --- p.105, Chapter 4.3 --- Characterization of ARHGEF 11 Promoter Region --- p.107, Chapter 4.3.1 --- Cloning of ARHGEF 11 Promoter --- p.107, Chapter 4.3.2 --- Promoter Activitiy of ARHGEFll Promoter --- p.109, Chapter 4.3.3 --- Deletion Mapping of ARHGEFll Promoter --- p.111, Chapter 4.3.4 --- Promoter Activities and Inducibitiy of Truncated ARHGEF 11 Promoters --- p.113, Chapter 4.4 --- Identifying Transcription Factor Binding Sites on ARHGEF 11 Promoter --- p.115, Chapter 4.4.1 --- Computational Prediction of Transcription Factor Binding Sites onARHGEFll Promoter --- p.115, Chapter 4.4.2 --- Preparation of Probe and Specific Competitors for EMSA --- p.117, Chapter 4.4.3 --- Interaction between DU145 Nuclear Extract and ARHGEF 11 Promoter Region --- p.119, Chapter 4.5 --- Biological Significances of miR-765 --- p.122, Chapter 4.2.1 --- "Effects of ICI 182,780 on DU145 Cell growth" --- p.122, Chapter 4.2.2 --- "Effects of ICI 182,780 on DU145 Cell Migration" --- p.124, Chapter 4.2.3 --- Verifying Functionality of Ectopic miR-765 --- p.131, Chapter 4.2.4 --- Effects of miR-765 on DU145 Cell Growth --- p.133, Chapter 4.2.5 --- Effects of miR-765 on DU145 Cell Migration --- p.135, Chapter Chapter 5: --- Discussion --- p.138, Chapter 5.1 --- "Identifying miR-765 as an Up-regulated miRNA by ICI 182,780" --- p.139, Chapter 5.1.1 --- "Information about ICI 182,780" --- p.139, Chapter 5.1.2 --- miRNA Profiling of DU145 --- p.139, Chapter 5.1.3 --- "Confirming Induction of miR-765 by ICI 182,780 and ERβ dependency with qRT-PCR" --- p.140, Chapter 5.1.4 --- "Up-regulation of miR-765 Host Gene, ARHGEF11, by ICI" --- p.141, Chapter 5.2 --- Regulatory Elements of miR-765 Expression --- p.143, Chapter 5.2.1 --- Own Upstream promoter of miR- 765 --- p.144, Chapter 5.2.2 --- Promoter of Host Gene ARHGEF11 --- p.146, Chapter 5.2.3 --- Interaction between ARHGEF11 Promoter Critical Region and Transcription Factors --- p.147, Chapter 5.2.4 --- Involvement of independent Promoter and Host Gene Promoter in miR-765 Regulation --- p.757, Chapter 5.3 --- Biological Significances of miR-765 on DU145 --- p.153, Chapter 5.4 --- Significance of Findings and Future Studies --- p.158, Chapter 5.4.1 --- Clinical Significance --- p.158, Chapter 5.4.2 --- Future Studies --- p.161, Chapter Chapter 6: --- Conclusion --- p.163, Chapter Chapter 7: --- References --- p.166, http://library.cuhk.edu.hk/record=b5894715, Use 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/)

Xu, Dandan., Thesis (Ph.D.)--Chinese University of Hong Kong, 2011., Includes bibliographical references (leaves 209-231)., Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web., also in Chinese., http://library.cuhk.edu.hk/record=b6075367, Use 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/)

In recent years, three dimensional (3D)-culture technique has emerged as a very popular approach to reconstruct tissue architectures and develop experimental models for studying epithelial cancers. However, 3D culture models of prostate epithelial cells to mimic prostate cancer development are relatively rare, making it highly desirable to develop and characterize novel 3D culture models suitable for studying prostate cancer. Recently, epithelial-mesenchymal transition (EMT) has emerged as an important mechanism for cancer cell invasion. The zinc finger transcriptional factor Snail as a key regulator of EMT has been found to contribute to aggressive progression in many types of neoplasms. Even though several studies corroborated that EMT is implicated in prostate cancer, the expression patterns of Snail in normal prostate and prostate cancer, and the functional role of Snail in prostate cancer as well as its relation with EMT are still unknown. Based on this background, my major efforts were to establish a 3D culture model of human prostatic epithelial cells with structural and functional relevance to prostate gland and to employ this model to study the functional role of Snail in the prostate cancer., When embedded in Matrigel for 3D culture, BPH-1 cells developed into growth-arrested acinar structures with a hollow lumen. Ultrastructural examination of BPH-1 spheroids by electricon microscopy indicated that BPH-1 spheroids displayed a polarized differentiation phenotype. Immunoflurescence analysis of polarized epithelial markers further confirmed that BPH-1 spheroids were polarized. In contrast, tumorigenic BPH-1CAFTD cells exhibited disorganized and continuously proliferating structures in Matrigel, with polarized epithelial markers randomly diffused or completely lost. In addition, BPH-1 CAFTD cells displayed significantly higher invasive capacity in comparison to BPH-1 cells by transwell invasion assay. Moreover, LY294002 treatment of BPH-1CAFTD1 and BPH-1CAFTD3 cells in 3D cultures resulted in impaired cell proliferation as evidenced by reduced colony size and decreased Ki-67 index, and western blot analysis showed that cyclin D1 protein levels were significantly decreased, while p21 protein levels were slightly up-regulated in LY294002-treated 3D cultures. Additionally, LY94002 significantly decreased the invasive capacity of BPH-1CAFTD1 and BPH-1CAFTD3 cells. Interestingly, LY294002 treatment completely reverted the disorganized non-polar 3D structures of BPH-1CAFTD1 cells to well-organized polarized spheroid structures in Matrigel, but failed to restore the polarized differentiation in 3D cultures of BPH-1CAFTD3 cells, which still formed compact aggregates as shown by confocal immunofluorescence analysis. Snail protein was barely detected in the epithelial cells of human benign prostatic tissue but significantly elevated as nuclear protein in primary prostate cancer and bone metastatic specimens by immunohistochemical analysis. Snail transcript levels were weakly expressed in a majority of nonmalignant prostatic epithelial cell lines, while markedly increased in almost all tested cancer cell lines. Snail expression induced a morphological switch to more s, Chu, Jianhong., Adviser: Franky Chan Leung., Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3448., Thesis (Ph.D.)--Chinese University of Hong Kong, 2008., Includes bibliographical references (leaves 143-166)., 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., s in English and Chinese., School code: 1307., isbn: 9781109225037, Use 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/)