Specific expression and androgen regulation of prostatic secretory protein of 94 amino acids (PSP94) in rat prostate gland.

• Other Authors: Kwong, Joseph.
• Format: Others
• Language: English; Chinese
• Published: 1999
• Subjects: Prostatic Neoplasms > genetics; Proteins > genetics; Prostate
• Online Access: http://library.cuhk.edu.hk/record=b5896335; http://repository.lib.cuhk.edu.hk/en/item/cuhk-322873

by Kwong Joseph. === Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. === Includes bibliographical references (leaves 142-164). === Abstracts in English and Chinese. === Abstract --- p.i === Acknowledgements --- p.iv === Abbreviations --- p.v === Table of contents --- p.vi === Chapter Chapter 1 --- Introduction === Chapter 1.1 --- Prostatic Secretory Proteins --- p.1 === Chapter 1.2 --- Rat Prostatic Secretory Proteins --- p.1 === Chapter 1.2.1 --- Prostatic Secretory Proteins in Ventral Prostate --- p.2 === Chapter 1.2.1.1 --- Prostatic Binding Protein (PBP) --- p.2 === Chapter 1.2.1.2 --- Androgen-Suppressed Proteins of Rat Ventral Prostate --- p.6 === Chapter 1.2.1.3 --- The 20-kDa Protein --- p.8 === Chapter 1.2.1.4 --- Spermine-Binding Proteins --- p.9 === Chapter 1.2.1.5 --- Prostatic Acid Phosphatase (PAP) --- p.10 === Chapter 1.2.2 --- Prostatic Secretory Proteins in Dorsal Prostate --- p.12 === Chapter 1.2.2.1 --- Dorsal Proteins I and II (DP I and DPII) --- p.12 === Chapter 1.2.2.2 --- Seminal Vesicle Secretion II (SVSII) --- p.14 === Chapter 1.2.2.3 --- Probasin --- p.16 === Chapter 1.2.3 --- Prostatic Secretory Proteins in Lateral Prostate --- p.18 === Chapter 1.3 --- Human Prostatic Secretion --- p.18 === Chapter 1.4 --- Human Prostatic Secretory Proteins --- p.18 === Chapter 1.4.1 --- Prostatic Acid Phosphatase (PAP) --- p.19 === Chapter 1.4.2 --- Prostate Specific Antigen (PSA) --- p.22 === Chapter 1.4.2.1 --- Molecular Biology of PSA --- p.22 === Chapter 1.4.2.2 --- Synthesis of PSA --- p.23 === Chapter 1.4.2.3 --- Kallikrein Gene Family --- p.23 === Chapter 1.4.2.4 --- Physiological Function of PSA --- p.24 === Chapter 1.4.2.5 --- PSA as an Immunohistochemical Marker --- p.25 === Chapter 1.4.2.6 --- PSA is not a Prostate-Specific Molecule --- p.26 === Chapter 1.4.3 --- Prostatic Secretory Protein of 94 Amino Acids (PSP94) --- p.27 === Chapter 1.4.3.1 --- Nucleotide Sequence of the PSP94 cDNA --- p.28 === Chapter 1.4.3.2 --- Amino Acid sequence of PSP94 --- p.28 === Chapter 1.4.3.3 --- Biological Properties of PSP94 --- p.29 === Chapter 1.4.3.4 --- Physiological Roles of PSP94 --- p.31 === Chapter 1.4.3.5 --- PSP94 and Its mRNA in Other Non-Prostatic Tissue --- p.31 === Chapter 1.4.3.6 --- PSP94 as a Tumor Marker of Prostate Cancer --- p.32 === Chapter 1.4.3.7 --- Homologous Proteins of PSP94 --- p.34 === Chapter 1.5 --- Aim of Study --- p.35 === Chapter Chapter 2 --- Materials and Methods === Chapter 2.1 --- Origin and Supply of Noble Rat --- p.37 === Chapter 2.2 --- Chemicals --- p.37 === Chapter 2.3 --- Bilateral Ochidectomy of Animals --- p.37 === Chapter 2.4 --- Androgen Replacement --- p.38 === Chapter 2.5 --- Hormonal and Drug Treatments on Castrated Animals --- p.38 === Chapter 2.6 --- Induction of Prostatic Intraepithelial Neoplasia in Noble Rat Prostate Gland by Long-Term Treatment with Steroids --- p.39 === Chapter 2.6.1 --- Preparation of Steroid Hormone-Filled Silastic® Tubings --- p.39 === Chapter 2.6.2 --- Surgical Implantation of Silastic® Tubings --- p.39 === Chapter 2.6.3 --- Protocols of Hormonal Treatments --- p.40 === Chapter 2.7 --- Androgen-Dependent Rat Dunning Prostatic Adenocarcinoma --- p.40 === Chapter 2.8 --- Androgen-Independent Prostatic Carcinoma Line (AIT) of Noble Rat --- p.41 === Chapter 2.9 --- Plasmids --- p.41 === Chapter 2.10 --- Restriction Enzyme Digestions of pLvB10 and cM-403 --- p.42 === Chapter 2.11 --- Amplification of Rat SVSII cDNA Fragment by RT-PCR and Subcloning --- p.42 === Chapter 2.12 --- Purification of DNA Fragment from Agarose Gel --- p.43 === Chapter 2.13 --- Subcloning of DNA into Vector --- p.44 === Chapter 2.14 --- Tissue Preparation for In-situ Hybridization --- p.47 === Chapter 2.15 --- Synthesis of Digoxigenin (DIG)-Labeled RNA Probe --- p.47 === Chapter 2.16 --- In-situ Hybridization --- p.48 === Chapter 2.17 --- Total RNA Extraction --- p.50 === Chapter 2.18 --- Northern Blotting Analysis --- p.51 === Chapter 2.19 --- Primers and Cycling Conditions --- p.53 === Chapter 2.20 --- Reverse Transcription Polymerase Chain Reaction (RT-PCR) --- p.54 === Chapter 2.21 --- Southern Blotting Analysis --- p.56 === Chapter 2.21.1 --- Southern Blotting --- p.56 === Chapter 2.21.2 --- Preparation of DIG-dUTP Labeled Rat PSP94 cDNA Probe --- p.56 === Chapter 2.21.3 --- Hybridization --- p.57 === Chapter 2.22 --- Restriction Mapping --- p.58 === Chapter 2.23 --- Semi-Quantitative RT-PCR --- p.59 === Chapter 2.24 --- Statistical Analysis --- p.59 === Chapter 2.25 --- "Protein Extraction, SDS-PAGE and Western Blotting Analysis" --- p.60 === Chapter 2.26 --- Immunohistochemistry --- p.63 === Chapter Chapter 3 --- Results === Chapter 3.1 --- Subcloning of DNAs into Vector --- p.65 === Chapter 3.1.1 --- Subcloning of 18s Ribosomal RNA cDNA Fragment --- p.65 === Chapter 3.1.2 --- Subcloning of Probasin cDNA Fragment --- p.66 === Chapter 3.1.3 --- Subcloning of SVSII cDNA Fragment --- p.66 === Chapter 3.1.4 --- Restriction Enzyme Mapping for PCR Product of SVSII --- p.67 === Chapter 3.2 --- Detection of mRNA and Protein Expression of PSP94 in Normal Rat Prostates --- p.68 === Chapter 3.2.1 --- In-situ Hybridization --- p.68 === Chapter 3.2.2 --- Northern Blotting --- p.68 === Chapter 3.2.3 --- RT-PCR Amplification --- p.69 === Chapter 3.2.4 --- Immunohistochemistry --- p.69 === Chapter 3.2.5 --- Western Blotting --- p.70 === Chapter 3.3 --- Detection of mRNA Expression of Probasin and SVSII in Normal Rat Prostates --- p.71 === Chapter 3.3.1 --- In-situ Hybridization --- p.71 === Chapter 3.3.2 --- RT-PCR Amplification --- p.71 === Chapter 3.4 --- "Androgen Regulation of PSP94,Probasin and SVSII mRNA Expression" --- p.72 === Chapter 3.4.1 --- In-situ Hybridization --- p.72 === Chapter 3.4.2 --- "Relative Expression Levels of PSP94, Probasin and SVSII mRNA in Normal, Castrated and Androgen Replaced Rat Lateral Prostates as Measured by a Semiquantitative RT-PCR Method" --- p.73 === Chapter 3.4.2.1 --- Determination of Exponential Range of PCR --- p.73 === Chapter 3.4.2.2 --- Semi-Quantitative RT-PCR --- p.74 === Chapter 3.4.3 --- Western Blot Analysis --- p.75 === Chapter 3.5 --- "Effect of Steroid Hormones and Zinc on the PSP94, Probasin and SVSII Expressions in Castrated Rat Prostates" --- p.76 === Chapter 3.5.1 --- Semi-Quantitative RT-PCR --- p.76 === Chapter 3.5.2 --- Western Blot Analysis --- p.77 === Chapter 3.6 --- "Detection of PSP94, Probasin and SVSII mRNA Expression in Dysplastic and Neoplastic Rat Prostates" --- p.78 === Chapter 3.6.1 --- "Detection of PSP94, Probasin and SVSII mRNA Expression in T+E2-Induced Prostatic Intraepithelial Neoplasia (PIN) of the Lateral Prostate of Noble Rats by In-situ Hybridization" --- p.78 === Chapter 3.6.2 --- "Detection of PSP94,Probasin and SVSII mRNA Expression in Dunning Tumor and AIT Prostatic Tumor" --- p.79 === Chapter 3.6.2.1 --- In-situ Hybridization --- p.79 === Chapter 3.6.2.2 --- RT-PCR Amplification --- p.79 === Chapter Chapter 4 --- Discussion === Chapter 4.1 --- Specific Expression of PSP94 in the Lateral Lobe of Rat Prostate --- p.114 === Chapter 4.2 --- Androgen Regulation of PSP94 --- p.118 === Chapter 4.2.1 --- Molecular Mechanism of Androgen Action --- p.118 === Chapter 4.2.2 --- Androgen Regulation of PSP94 in Rat Lateral Prostate --- p.121 === Chapter 4.3 --- "Effect of Steroid Hormones and Zinc on the PSP94, Probasin and SVSII Expressions in Castrated Rat Lateral Prostate" --- p.126 === Chapter 4.4 --- "Detection of PSP94, Probasin, SVSII mRNA Expression in Dysplastic and Neoplastic Rat Prostates" --- p.133 === Chapter 4.5 --- Gene Therapy --- p.139 === Chapter Chapter 5 --- Conclusions --- p.141 === References --- p.142 === Appendixes --- p.165