Chan Chun-pong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 171-176). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 摘要 --- p.iii / Abbreviation --- p.iv / Table of contents --- p.v / List of figures --- p.xi / List of tables --- p.xvi / Chapter Chapter 1 --- introduction / Chapter 1.1 --- Traditional Chinese medicines (TCMs) --- p.2 / Chapter 1.2 --- Liver disorders in Asia Pacific region --- p.3 / Chapter 1.3 --- Classification of liver disorders --- p.7 / Chapter 1.3.1 --- Hepatitis --- p.8 / Chapter 1.3.1.1 --- Hepatitis A virus infection --- p.8 / Chapter 1.3.1.2 --- Hepatitis B virus infection --- p.9 / Chapter 1.3.1.3 --- Hepatitis C virus infection --- p.11 / Chapter 1.3.1.4 --- Hepatitis D virus infection --- p.12 / Chapter 1.3.1.5 --- Hepatitis E virus infection --- p.12 / Chapter 1.3.2 --- Cancer of the liver --- p.13 / Chapter 1.3.2.1 --- Hepatocellular carcinoma --- p.13 / Chapter 1.3.2.2 --- Cholangiocarcinoma --- p.14 / Chapter 1.3.2.3 --- Metastatic liver cancer --- p.14 / Chapter 1.4 --- Conventional treatment of liver disorders --- p.14 / Chapter 1.5 --- Role of traditional Chinese medicines in hepatoprotective functions --- p.16 / Chapter 1.5.1 --- Abri Herba (Abrus Cantoniensis Hance) --- p.17 / Chapter 1.5.2 --- Rhizoma Coptidis (Coptidis chinensis Franch) --- p.18 / Chapter 1.5.3 --- Fructus Forsythia (Forsythia suspense (Thunb) Vahl) --- p.22 / Chapter 1.6 --- Molecular studies of hepatoprotective effects of TCMs --- p.26 / Chapter 1.6.1 --- Roles of detoxofication enzymes in hepatoprotection --- p.27 / Chapter 1.6.2 --- Studies of growth-related genes in cell cycle control --- p.29 / Chapter 1.7 --- Aims of project --- p.32 / Chapter 1.8 --- Application of the project --- p.33 / Chapter Chapter 2 --- Methods and materials --- p.34 / Chapter 2.1 --- Screening of traditional Chinese medicines --- p.35 / Chapter 2.2 --- Preparation of TCMs --- p.35 / Chapter 2.2.1 --- Preparation of aqueous extracts of TCMs --- p.35 / Chapter 2.2.2 --- Preparation of active components of TCMs --- p.36 / Chapter 2.3 --- In vitro assays --- p.40 / Chapter 2.3.1 --- Cell culture --- p.40 / Chapter 2.3.2 --- Cytotoxicity test --- p.40 / Chapter 2.4 --- Screening of expressed gene induced by TCMs --- p.41 / Chapter 2.4.1 --- RNA preparation --- p.41 / Chapter 2.4.2 --- cDNA array hybridization --- p.42 / Chapter 2.4.3 --- Reverse Transcription --- p.43 / Chapter 2.5 --- Confirmation of expressed genes induced by TCMs --- p.44 / Chapter 2.5.1 --- Semi-quantitative PCR analysis --- p.44 / Chapter 2.5.2 --- Northern blot analysis --- p.46 / Chapter 2.6 --- Studies of effects of TCMs in gene expression --- p.47 / Chapter 2.6.1 --- Dosage-course study --- p.47 / Chapter 2.6.2 --- Time-course study --- p.48 / Chapter Chapter 3 --- Results --- p.50 / Chapter 3.1 --- "Cytotoxicity test of A.H., R.C. and F.F" --- p.51 / Chapter 3.2 --- "Molecular screening of expressed gene induced by A.H., R.C., F.F" --- p.58 / Chapter 3.3 --- Confirmation of expressed gene using semi-quantitative RT- PCR --- p.70 / Chapter 3.3.1 --- Dosage-course and time-course studies of A.H. using RT- PCR --- p.70 / Chapter 3.3.2 --- Dosage-course and time-course studies of R.C. using RT- PCR --- p.94 / Chapter 3.3.3 --- Dosage-course and time-course studies of A.H. using RT- PCR --- p.113 / Chapter 3.4 --- Confirmation of expressed gene using northern blot anaylsis --- p.118 / Chapter 3.4.1 --- Dosage-course and time-course studies of effects of A.H. and L- abrine in Northern blot analysis --- p.118 / Chapter 3.4.2 --- Dosage-course and time-course studies of effects of R.C. and berberine in Northern blot analysis --- p.129 / Chapter 3.4.3 --- Dosage-course and time-course studies of effects of F.Fin Northern blot analysis --- p.147 / Chapter Chapter 4 --- Discussion --- p.152 / Chapter 4.1 --- "Roles of A.H., R.C. and F.F. in treatment and prevention of liver disorders" --- p.153 / Chapter 4.2 --- "Cytotoxicity effect A.H., R.C., and F.F. in liver cells" --- p.153 / Chapter 4.3 --- Effects of herbal medicines on the transcription of mRNA in liver cells --- p.155 / Chapter 4.3.1 --- Effects of treatment of A.H. in liver at transcriptional level … --- p.155 / Chapter 4.3.2 --- Effects of treatment of R.C. in liver at transcriptional level … --- p.156 / Chapter 4.3.3 --- Effects of treatment of R.C. in liver at transcriptional level --- p.157 / Chapter 4.4 --- Comparison of results of RT-PCR and Northern blot analysis --- p.157 / Chapter 4.4.1 --- Comparison of the effects of time and dosage-course studies of DTD expression induced by A.H. and L-abrine --- p.157 / Chapter 4.4.2 --- Comparison of the effects of time and dosage-course studies of p21;cip;waf1 expression induced by A.H. and L-abrine --- p.158 / Chapter 4.4.3 --- Comparison of the effects of time and dosage-course studies of c-myc responsive protein; rcl expression induced by R.C. and berberine --- p.159 / Chapter 4.4.4 --- Comparison of the effects of time and dosage-course studies of GST Ya expression induced by R.C. and berberine --- p.160 / Chapter 4.4.5 --- Comparison of the effects of time and dosage-course studies of GST 7-7 expression induced by F.F --- p.160 / Chapter 4.5 --- Biochemical significance of genes induced by hepatoprotective TCMs --- p.161 / Chapter 4.5.1 --- Roles of significant expression of detoxifying enzymes induced by TCMs in liver cells --- p.161 / Chapter 4.5.2 --- Roles of induction of growth-related c-myc responsive protein; rcl in R.C. treated liver cells --- p.167 / Chapter 4.5.3 --- Roles of increased p21;cip;waf1 expression in A.H. treated liver cells --- p.168 / Chapter 4.6 --- Conclusion --- p.169

Wong, Wai Ki Vicky. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 105-131). / Abstracts in English and Chinese. / Abstract --- p.ii / 摘要 --- p.iv / Acknowledgements --- p.vi / Table of contents --- p.vii / List of tables --- p.xii / List of figures --- p.xiii / List of abbreviations --- p.xiv / Chapter Chapter 1: --- Introduction --- p.1 / Chapter Chapter 2: --- Colorectal cancer / Chapter 2.1 --- Literature Review / Chapter 2.1.1 --- Colorectal cancer --- p.8 / Chapter 2.1.2 --- Incident rate of colorectal cancer --- p.8 / Chapter 2.1.3 --- Hereditary colorectal cancer --- p.9 / Chapter 2.1.4 --- Sporadic colorectal cancer and Wnt signaling pathway --- p.10 / Chapter 2.1.5 --- Chemotherapy treatment of colorectal cancer --- p.11 / Chapter 2.1.5.1 --- 5-Fluorouracil --- p.12 / Chapter 2.1.5.2 --- Oxaliplatin --- p.14 / Chapter 2.1.5.3 --- Irinotecan --- p.14 / Chapter 2.1.6 --- Biomarkers for colorectal cancer --- p.15 / Chapter 2.1.6.1 --- Thymidylate synthase --- p.15 / Chapter 2.1.6.2 --- Dihydropyrimidine dehydrogenase --- p.16 / Chapter 2.1.6.3 --- Thymidine phosphorylase --- p.16 / Chapter 2.1.6.4 --- Microsatellite-instability status --- p.16 / Chapter 2.1.6.5 --- Clinical uses of biomarkers for colorectal cancer --- p.17 / Chapter 2.1.7 --- Choice of cell line as colorectal cancer model --- p.17 / Chapter 2.1.8 --- Aims of study --- p.17 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Verification of SW480 as a nuclear β-catenin positive cell line / Chapter 2.2.1.1 --- Maintenance of cell lines --- p.21 / Chapter 2.2.1.2 --- Antibody --- p.21 / Chapter 2.2.1.3 --- Agar block preparation for SW480 and CCD-18C0 cells --- p.22 / Chapter 2.2.1.4 --- Immunocytochemical staining --- p.22 / Chapter 2.2.2 --- Effect of anti-cancer drugs on cell viability / Chapter 2.2.2.1 --- Maintenance of cell lines --- p.22 / Chapter 2.2.2.2 --- MTT cell viability assay --- p.23 / Chapter 2.3 --- Results / Chapter 2.3.1 --- SW480 is a β-catenin positive cell line --- p.24 / Chapter 2.3.2 --- Antiproliferative effects of cytotoxic drugs in SW480 cells / Chapter 2.3.2.1 --- 5-Fluorouracil --- p.26 / Chapter 2.3.2.2 --- Oxaliplatin --- p.29 / Chapter 2.3.2.3 --- Irinotecan --- p.31 / Chapter 2.4 --- Discussion / Chapter 2.4.1 --- SW480 as a nuclear β-catenin positive cell line --- p.33 / Chapter 2.4.2 --- Antiproliferative effects of 5-fluorouracil in SW480 cells --- p.33 / Chapter 2.4.3 --- Summary --- p.34 / Chapter Chapter 3: --- Effect of 5-fluorouracil on mRNA expression in SW480 cells / Chapter 3.1 --- Literature Review / Chapter 3.1.1 --- Application of quantitative real-time polymerase chain reaction in cancer research / Chapter 3.1.1.1 --- Principles of quantitative real-time polymerase chain reaction --- p.36 / Chapter 3.1.1.2 --- Advantages of quantitative real-time polymerase chain reaction over conventional polymerase chain reaction --- p.39 / Chapter 3.1.1.3 --- Determination of colorectal cancer biomarkers by quantitative real-time polymerase chain reaction --- p.39 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Determination of the effect of 5-fluorouracil on mRNA expression in SW480 cells / Chapter 3.2.1.1 --- Treatment of cells --- p.40 / Chapter 3.2.1.2 --- Extraction of total RNA from SW480 cells --- p.40 / Chapter 3.2.1.3 --- Removal of genomic DNA --- p.41 / Chapter 3.2.1.4 --- Determination of the efficiency of genomic DNA removal --- p.42 / Chapter 3.2.1.5 --- Determination of the purity and concentration of RNA --- p.42 / Chapter 3.2.1.6 --- Determination of the integrity of RNA --- p.43 / Chapter 3.2.1.7 --- First strand cDNA synthesis --- p.44 / Chapter 3.2.1.8 --- Real-time polymerase chain reaction using human Wnt signaling pathway RT2 ProfileŕёØ PCR array --- p.44 / Chapter 3.2.1.9 --- Calculation of the fold-change in genes expression between the 5-FU treated and control SW480 cells --- p.45 / Chapter 3.3 --- Results / Chapter 3.3.1 --- The quality and quantity of RNA --- p.46 / Chapter 3.3.2 --- Effects of 5-fluorouracil on genes expression in SW480 cells --- p.48 / Chapter 3.4 --- Discussion / Chapter 3.4.1 --- Alterations in mRNA expression in 5-fluorouracil treated SW480 cells --- p.55 / Chapter 3.4.1.1 --- Extracellular signaling molecules --- p.55 / Chapter 3.4.1.2 --- Canonical Wnt signaling pathway --- p.56 / Chapter 3.4.1.3 --- Regulators of cell cycle --- p.57 / Chapter 3.4.1.4 --- Regulators of growth and proliferation --- p.58 / Chapter 3.4.1.5 --- Regulators of transcription --- p.58 / Chapter 3.4.1.6 --- Regulators of Wnt receptor signaling pathway --- p.60 / Chapter 3.4.1.7 --- Other genes involved in Wnt signaling --- p.61 / Chapter 3.4.2 --- Limitations of Q-RT-PCR --- p.61 / Chapter 3.4.3 --- Summary --- p.62 / Chapter Chapter 4: --- Effect of 5-fluorouracil on proteins expression in SW480 cells / Chapter 4.1 --- Literature Review / Chapter 4.1.1 --- From mRNA to proteins --- p.63 / Chapter 4.1.2 --- Application of proteomics in cancer research --- p.63 / Chapter 4.1.3 --- Two-dimensional gel electrophoresis --- p.64 / Chapter 4.1.4 --- Principles of MALDI TOF mass spectrometry --- p.64 / Chapter 4.1.5 --- Peptide mass fingerprinting --- p.65 / Chapter 4.1.6 --- Drug response proteins detected by proteomics in colorectal cancer cell lines --- p.65 / Chapter 4.1.7 --- Detection of biomarker in colorectal cancer formation using proteomics --- p.66 / Chapter 4.2 --- Materials and Methods / Chapter 4.2.1 --- Determination of the effect of 5-fluorouracil on proteins expression in SW480 cells / Chapter 4.2.1.1 --- Treatment of cells --- p.67 / Chapter 4.2.1.2 --- Cell lysis --- p.67 / Chapter 4.2.1.3 --- Protein quantitation of cell lysate --- p.67 / Chapter 4.2.1.4 --- Sample preparation for two-dimensional electrophoresis --- p.68 / Chapter 4.2.1.5 --- Two-dimensional electrophoresis --- p.69 / Chapter 4.2.1.6 --- Silver staining --- p.69 / Chapter 4.2.1.7 --- Image analysis --- p.70 / Chapter 4.2.1.8 --- In-gel protein digestion --- p.70 / Chapter 4.2.1.9 --- Peptide mass fingerprinting using mass spectrometry --- p.71 / Chapter 4.3 --- Results / Chapter 4.3.1 --- Protein expression patterns of 5-fluorouracil treated and untreated SW480 cells by 2-dimensional electrophoresis --- p.72 / Chapter 4.3.2 --- Identification of the differentially expressed proteins after 5-fluorouracil treatment in SW480 cells --- p.75 / Chapter 4.4 --- Discussion / Chapter 4.4.1 --- Effects of 5-fluorouracil on protein expression in SW480 cells --- p.82 / Chapter 4.4.1.1 --- Identified upregulated proteins after 5-fluorouracil treatment in SW480 cells / Chapter 4.4.1.1.1 --- Cyclophilin A --- p.83 / Chapter 4.4.1.1.2 --- Cytokeratin 19 --- p.83 / Chapter 4.4.1.1.3 --- Cytokeratin 8 --- p.84 / Chapter 4.4.1.1.4 --- RAN --- p.84 / Chapter 4.4.1.1.5 --- Heat shock protein 27 --- p.84 / Chapter 4.4.1.1.6 --- Peroxiredoxin 6 --- p.85 / Chapter 4.4.1.2 --- Identified dowiiregulated proteins after 5-fluorouracil treatment in SW480 cells / Chapter 4.4.1.2.1 --- Heat shock protein 60 --- p.86 / Chapter 4.4.1.2.2 --- Cytokeratin 18 --- p.86 / Chapter 4.4.1.2.3 --- Cytokeratin 9 --- p.86 / Chapter 4.4.1.2.4 --- Carbamoylphosphate synthetase I --- p.87 / Chapter 4.4.1.2.5 --- a-Enolase --- p.87 / Chapter 4.4.1.2.6 --- Heat shock protein 70 --- p.87 / Chapter 4.4.1.2.7 --- nm23 --- p.88 / Chapter 4.4.1.2.8 --- β-actin --- p.88 / Chapter 4.4.2 --- Limitations of proteomics profiling --- p.89 / Chapter 4.4.3 --- Summary --- p.90 / Chapter Chapter 5: --- Verification of proteinśة identities by immunocytochemical staining / Chapter 5.1 --- Materials and Methods / Chapter 5.1.1 --- Antibodies --- p.91 / Chapter 5.1.2 --- Treatment of cells --- p.91 / Chapter 5.1.3 --- Agar block preparation of SW480 cells --- p.92 / Chapter 5.1.4 --- Immunocytochemical staining and evaluation --- p.92 / Chapter 5.1.5 --- Polymer-based immunohistochemical detection system --- p.93 / Chapter 5.1.6 --- Statistical analyses --- p.93 / Chapter 5.2 --- Results / Chapter 5.2.1 --- Confirmation of proteomic findings using immunocytochemical stainings in paraffin-embedded sections of 5-fluorouracil treated and untreated SW480 cells --- p.94 / Chapter 5.3 --- Discussion / Chapter 5.3.1 --- Immunocytochemical staining to verify proteomics findings of 5-fluorouracil treated and untreated SW480 cells --- p.99 / Chapter 5.3.2 --- Limitations of ICC staining --- p.100 / Chapter 5.3.3 --- Summary --- p.100 / Chapter Chapter 6: --- Conclusions and future perspectives / Chapter 6.1 --- Significance of study --- p.101 / Chapter 6.2 --- Future perspectives --- p.102 / References --- p.105