Diss. (sammanfattning) Stockholm : Karolinska institutet, 2005. / Härtill 5 uppsatser.

Aim. The aim of this study was to synthesize and characterize novel analogues of [DACH-Pt-DMC] by using different stereoisomers of DACH; and to investigate any differences in in vitro activity of these complexes in human hepatocellular carcinoma (HCC), colorectal carcinoma (CRC) cell lines and acquired cisplatin or oxaliplatin resistant sub-lines, and to compare that of oxaliplatin and other established Pt-based anticancer agents. Mechanistic roles of DACH-Pt- and DMC components of the TCM-Pt complexes on affecting HCT 116 human CRC cell line were investigated by flow cytometry, COMET assay and cDNA microarray analysis. / Background. Demethylcantharidin (DMC), a modified component of the traditional Chinese medicine (TCM), integrated with a platinum (Pt) moiety created a series of TCM-Pt complexes [Pt(C8H8O 5)(NH2R)2] 1-5 which demonstrated superior antitumor activity and circumvention of cisplatin resistance in vitro. Compound 5, derived from the 1,2-diaminocyclohexane (DACH) ligand (where R=trans-C6H10) had the most potent antitumor activity and closest structural resemblance to oxaliplatin (R,R-DACH-Pt complex) which is the first Pt-based anticancer drug to demonstrate convincing clinical activity against colorectal cancer and has a mechanism of action and resistance that is clearly different from that of cisplatin and carboplatin. / Conclusion. This study is the first to examine the mechanism of anticancer activity of new complexes that integrate DMC with different isomers of DACH. It has shown that both DACH-Pt- and DMC components contribute significantly to the compounds' potent anticancer activity, but likely with different mechanisms of action. The DACH-Pt- component appears to dictate the cell cycle distribution, whereas the DMC component appears to enhance cytotoxicity by inducing more DNA damage in HCT 116 colorectal cancer cells. / Methods. DMC was reacted with appropriate DACH-Pt-(NO3) 2 intermediates, which were prepared from treatment of K2PtCl 4 with stereoisomeric DACH (RR-, SS- & cis-), followed by reaction with silver nitrate. Proton NMR, high-resolution MS, polarimetry and circular dichroism (CD) spectroscopy were used to characterize their chemical structures and optical activities. In vitro antitumor activity (IC50 of 72hr drug exposure time) were assessed by a standard MTT assay. Cell cycle analysis by flow cytometry was determined at 0, 6, 12, 18, 24, 48 and 72 h after drug treatment (cisplatin, carboplatin, oxaliplatin, DMC, compound 1 or trans-DACH-Pt-DMC analogues) at IC50 and 5 x IC50 concentrations with three to four replicates. Comet assay was performed with a fluorescent microscope and used to examine DNA damage after drug treatments (50muM of cisplatin, carboplatin, oxaliplatin, DMC, compound 1 or R,R-DACH-Pt-DMC) for 3hr. cDNA microarray was performed on Affymetrix Human Genome U133A Set and used to analyze gene expression profiles in HCT 116 exposed to trans-(+/-)-DACH-Pt-DMC or oxaliplatin at their IC50 for 72hr. / Results. The in vitro results showed that the trans-analogues were consistently the most potent amongst all the compounds tested in both HCC and CRC cell lines: the trans-(+)(1R,2R)-DACH-Pt-DMC complex, in particular, was the most effective stereoisomer. All of the stereoisomeric DACH-Pt-DMC complexes and oxaliplatin were apparently able to circumvent cisplatin resistance in Huh-7 and SK-Hep1 sub-lines, but cross resistant with oxaliplatin in HCT 116 oxaliplatin resistant sub-line. Flow cytometric analysis revealed the novel trans-DACH-Pt-DMC analogues and oxaliplatin behaved similarly: that is, the compounds at 5 x IC50 concentrations all caused a significant decrease in the S-phase population within 18h and at the same time induced G2/M arrest, and without obvious sub-G 1 phase accumulation, but distinct from that of cisplatin, carboplatin or DMC. Comet assay showed that trans-(+)-(1R,2 R)-DACH-Pt-DMC caused the most significant DNA damage at an equivalent molar concentration. Microarray analysis suggested that the mechanistic role of the DMC ligand can induce the cell cycle to accelerate from the G 1 to S-phase and cause M-phase arrest. / Yu Chun Wing. / "July 2006." / Advisers: Yee-ping Ho; Chik Fun Steve Au-Yeung. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1586. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 191-232). / 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. / Abstracts in English and Chinese. / School code: 1307.

Leung Hau Yi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 116-131). / Abstracts in English and Chinese. / Thesis Committee --- p.in / Acknowledgement --- p.II / Abstract --- p.III / 摘要 --- p.V / Table of Contents --- p.VII / List of Figures --- p.X / List of Tables --- p.XIII / Appendix Abbreviations --- p.XIV / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Breast Cancer --- p.1 / Chapter 1.2 --- Development of Breast Cancer --- p.2 / Chapter 1.3 --- Risk Factors of Breast Cancer --- p.3 / Chapter 1.3.1 --- Age --- p.3 / Chapter 1.3.2 --- Genetic Factors --- p.4 / Chapter 1.3.3 --- Hormonal Factors --- p.5 / Chapter 1.3.4 --- Lifestyles --- p.6 / Chapter 1.4 --- Drug Metabolism --- p.6 / Chapter 1.5 --- UGT1A1 --- p.7 / Chapter 1.5.1 --- UDP-glucuronosyltransferase --- p.7 / Chapter 1.5.2 --- UGT1A1 --- p.9 / Chapter 1.6 --- Cytochrome P450 I Enzyme Family --- p.10 / Chapter 1.6.1 --- CYP450 subfamily --- p.10 / Chapter 1.6.2 --- CYP1A1 --- p.11 / Chapter 1.6.3 --- CYP1B1 --- p.12 / Chapter 1.7 --- Reasons why UGT1A1 is being studied --- p.13 / Chapter 1.8 --- Outline of this Study --- p.14 / Chapter 1.8.1 --- Effects of Over-expressing UDP-Glucuronpsyltransferase and Cytochrome P450 1A1 Against Xenobiotic Assault in Breast Cancer Cells --- p.14 / Chapter 1.8.2 --- Effects of Genistein and Resveratrol on Phase I and II Enzymes in a Non-cancerous Breast Cell Line --- p.15 / Chapter 1.8.3 --- Effects of UGT1A1 on Cancer Drug Treatment --- p.15 / Chapter Chapter 2 --- Materials and Methods --- p.16 / Chapter 2.1 --- Chemicals --- p.16 / Chapter 2.2 --- Cell Culture --- p.16 / Chapter 2.2.1 --- Maintenance --- p.16 / Chapter 2.2.2 --- Preparation of Cell Stock --- p.17 / Chapter 2.2.3 --- Cell Recovery from Liquid Nitrogen Stock --- p.17 / Chapter 2.3 --- Cloning and Transfection --- p.18 / Chapter 2.3.1 --- Isolation of RNA from cells and cDNA synthesis --- p.18 / Chapter 2.3.2 --- Amplification of UGTlAl --- p.20 / Chapter 2.3.3 --- Separation and Purification of DNA from Agarose Gel --- p.21 / Chapter 2.3.4 --- Restriction Digestion --- p.22 / Chapter 2.3.5 --- Ligation of DNA Fragment and Vector --- p.22 / Chapter 2.3.6 --- Transformation of DH5a --- p.23 / Chapter 2.3.7 --- Small Scale Plasmid Purification (Miniprep) --- p.24 / Chapter 2.3.8 --- Large Scale Plasmid Purification (Maxiprep) --- p.25 / Chapter 2.3.9 --- Stable Transfection into MCF-7 cells with LipofectAMINE PLUS reagent --- p.26 / Chapter 2.4 --- Analytical Procedures --- p.27 / Chapter 2.4.1 --- Western Blot Analysis --- p.27 / Chapter 2.4.2 --- Measurement of cell proliferation (MTT assay) --- p.28 / Chapter 2.4.3 --- Measurement of DMBA-DNA Adduct Formation --- p.28 / Chapter 2.4.4 --- Comet Assay --- p.29 / Chapter 2.4.5 --- Relative Quantitative Real Time PCR --- p.30 / Chapter 2.4.5.1 --- Real Time PCR Using TaqMan Probe --- p.30 / Chapter 2.4.5.2 --- Statistical Analysis of 2-ΔΔCT Comparative Gene Expression --- p.31 / Chapter 2.4.6 --- Flow Cytometry --- p.31 / Chapter 2.4.7 --- EROD Activity in Intact Cells --- p.31 / Chapter 2.4.8 --- High Performance Liquid Chromatography --- p.32 / Chapter 2.5 --- Statistical Analysis --- p.34 / Chapter Chapter 3 --- Effects of Over-Expressing UDP-GIucuronosyltransferase and Cytochrome P450 1A1 Against Xenobiotic Assault in Breast Cancer Cells --- p.35 / Chapter 3.1 --- Introduction --- p.35 / Chapter 3.2 --- Results --- p.38 / Chapter 3.2.1 --- Effectiveness of Transfection --- p.38 / Chapter 3.2.2 --- Cell Proliferation Experiments --- p.41 / Chapter 3.2.3 --- Regulation of Estrogen Receptor (ER) Expression --- p.43 / Chapter 3.2.4 --- Formation of DMBA-DNA adduct formation --- p.45 / Chapter 3.2.5 --- Single Cell Gel Electrophoresis (Comet Assay) of DMBA-induced DNA Damage in MCF-7UGT1A1 cells --- p.46 / Chapter 3.2.6 --- HPLC for Estradiol-glucuronidation Analysis --- p.49 / Chapter 3.2.7 --- Single Cell Gel Electrophoresis (Comet Assay) of DMBA or E2-induced DNA Damage in MCF-7cyp1A1 cells --- p.51 / Chapter 3.3 --- Discussion --- p.56 / Chapter Chapter 4 --- Effects of Genistein and Resveratrol on Phase I and II Enzymes in a Non-Cancerous Breast Cell Line --- p.61 / Chapter 4.1 --- Introduction --- p.61 / Chapter 4.2 --- Results --- p.66 / Chapter 4.2.1 --- "Genistein and Resveratrol Reduced DMBA-induced UGT1A1, CYP1A1 and CYP1B1 Expression" --- p.66 / Chapter 4.2.2 --- Genistein and Resveratrol Reduced the Formation of DMBA-DNA Adduct in MCF-10A Cells --- p.73 / Chapter 4.2.3 --- Genistein and Resveratrol Reduced the Single Strand DNA Damage Generated by DMBA in MCF-10A Cells --- p.76 / Chapter 4.2.4 --- Genistein and Resveratrol Reduced DMBA-induced EROD Activities --- p.81 / Chapter 4.3 --- Discussion --- p.84 / Chapter Chapter 5 --- Effects of Ugtlal on Cancer Drug Treatment --- p.89 / Chapter 5.1 --- Introduction --- p.89 / Chapter 5.2 --- Results --- p.93 / Chapter 5.2.1 --- Cell Proliferation Experiment --- p.93 / Chapter 5.2.2 --- "Expression of Bcl-2 and Bax proteins in Paclitaxel- or VCR-treated MCF-7, MCF-7control and MCF-7UGt1A1 cells" --- p.98 / Chapter 5.2.3 --- Flow Cytometric Analysis of Cell Cycle Phase Distributionin Paclitaxel- or VCR-treated MCF-7 cells --- p.103 / Chapter 5.3 --- Discussion --- p.110 / Chapter Chapter 6 --- Summary --- p.114 / Bibliography --- p.116

Leung Chung Wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 142-146). / Abstracts in English and Chinese. / ABSTRACT (ENGISH) --- p.iii / ABSTRACT (CHINESS) --- p.v / ACHKNOWLEDGEMENTS --- p.vii / TABLE OF CONTENTS --- p.viii / Chapter CHAPTER 1 --- Introduction and Background / Chapter 1.1 --- Introduction of Platinum Drugs --- p.1 / Chapter 1.2 --- Mechanism of Action of Cisplatin --- p.3 / Chapter 1.3 --- Structure-Activity Relationships of the Platinum Drug 、 --- p.4 / Chapter 1.4 --- QS AR Parameters --- p.9 / Chapter 1.4.1 --- Chemical Hardness: Descriptor of Chemical Reactivity --- p.9 / Chapter 1.4.2 --- Possible Reaction Pathway of Platinum Drugs --- p.12 / Chapter 1.4.2.1 --- Proposed DNA Binding Pathway of Platinum Drugs --- p.13 / Chapter 1.4.2.1.1 --- Hydrolysis Pathway --- p.13 / Chapter 1.4.2.1.2 --- DNA Binding Pathway Involving the S-containing Biomolecules (Methionine Pathways) --- p.16 / Chapter 1.4.2.1.3 --- Conclusion --- p.21 / Chapter 1.5 --- Thesis Scope --- p.22 / Chapter CHAPTER 2 --- Theory and Methodology / Chapter 2.1 --- Introduction --- p.24 / Chapter 2.2 --- Density Functional Theory (DFT) --- p.24 / Chapter 2.2.1 --- Kohn-Sham Theorem --- p.25 / Chapter 2.2.2 --- Exchange-Correlation Energy Functional --- p.27 / Chapter 2.3 --- Basis Set --- p.27 / Chapter 2.3.1 --- Relativistic Effective Core Potential --- p.27 / Chapter 2.3.2 --- Double-Zeta --- p.28 / Chapter 2.3.3 --- Polarized Basis Set --- p.29 / Chapter 2.4 --- Solvation Model --- p.30 / Chapter 2.4.1 --- Continuum Model --- p.30 / Chapter 2.4.1.1 --- Simple Solvation Model --- p.31 / Chapter 2.4.1.1.1 --- Electrostatic Component --- p.31 / Chapter 2.4.1.1.2 --- Dispersion-Repulsion Interaction --- p.33 / Chapter 2.4.1.1.3 --- Cavitatoin Energy --- p.35 / Chapter 2.4.1.2 --- Polarized Continuum Model --- p.36 / Chapter 2.5 --- Methodology --- p.39 / Chapter 2.5.1 --- Calculation of DFT Global Reactivity Index --- p.39 / Chapter 2.5.1.1 --- Calculation for the Reaction Intermediates --- p.41 / Chapter 2.5.2 --- Calculation of the Reaction Pathways --- p.42 / Chapter CHAPTER 3 --- Results and Discussion / Chapter 3.1 --- Introduction --- p.49 / Chapter 3.2 --- Optimized Structure against Experimental Geometry --- p.49 / Chapter 3.3 --- Kohn-Sham Orbitals --- p.54 / Chapter 3.3.1 --- Location of the HOMO and LUMO --- p.55 / Chapter 3.4 --- Results of the DFT Reactivity Parameter --- p.57 / Chapter 3.5 --- Chemical Structure of the Drugs in the QSAR --- p.64 / Chapter 3.6 --- QSAR Analysis --- p.67 / Chapter 3.6.1 --- The Overall QSAR Plot of the Platinum Drugs --- p.68 / Chapter 3.6.1.1 --- Empirical Applicability of the QSAR on the Platinum(IV) Drugs --- p.70 / Chapter 3.6.1.2 --- Detail QASR Study According to the Type of Platinum Drug --- p.71 / Chapter 3.6.1.2.1 --- QSAR Study of the non-“trans-DACH´ح Platinum Drugs --- p.72 / Chapter 3.6.1.2.1.1 --- "QSAR Equation of the non-""trαns-DACH"" Platinum Drugs" --- p.75 / Chapter 3.6.1.2.2 --- QSAR Analysis for the Pt-trαns-DACH Drugs --- p.77 / Chapter 3.6.1.2.2.1 --- "QSAR Study of trans-S,S-DACH Platinum Drugs" --- p.79 / Chapter 3.6.1.2.2.2 --- "QSAR Study of trans-R,R-DACH Platinum Drugs" --- p.80 / Chapter 3.6.1.3 --- Summary --- p.81 / Chapter 3.7 --- QSAR Study of the Important Intermediates Using Chemical Hardness --- p.82 / Chapter 3.7.1 --- Optimized Structure for the Intermediates --- p.84 / Chapter 3.7.2 --- QSAR of the Dichloride Pt-Drugs Using Chemical Hardness of Parent Compounds --- p.90 / Chapter 3.7.3 --- QSAR of the Dichloride Pt-Drugs Using Chemical Hardness of Hydrolysis Intermediates --- p.91 / Chapter 3.7.4 --- QSAR of the Dichloride Pt-Drugs Using Chemical Hardness of Cyclic-Methionine Intermediates --- p.93 / Chapter 3.7.5 --- Conclusion --- p.95 / Chapter CHAPTER 4 --- Results and Discussion / Chapter 4.1 --- Introduction --- p.96 / Chapter 4.2 --- Study Scheme --- p.97 / Chapter 4.3 --- Optimized Structures --- p.98 / Chapter 4.4 --- Comments on the Reliability of the Calculation Model --- p.103 / Chapter 4.4.1 --- Reaction Profile in the Gas Phase --- p.104 / Chapter 4.4.2 --- Reaction Profiles Using Simple Solvation Model --- p.105 / Chapter 4.4.2.1 --- Defects of the Simple Solvation Model --- p.107 / Chapter 4.4.3 --- Reaction Profile Using PCM-UAHF Solvation Model --- p.109 / Chapter 4.4.3.1 --- Selection of the Reaction Parameters for the QSAR Study --- p.112 / Chapter 4.5 --- QSAR Study of Platinum Drugs Using the Reaction Parameters (AG and ΔG+) --- p.121 / Chapter 4.5.1 --- QSAR Analysis Using ΔG+(hydrolysis) --- p.121 / Chapter 4.5.2 --- QSAR Analysis Using ΔG(hydrolysis) --- p.123 / Chapter 4.5.3 --- QSAR Analysis Using ΔG+(guanine) --- p.125 / Chapter 4.5.4 --- QSAR Analysis Using ΔG(guanine) --- p.127 / Chapter 4.5.5 --- Further investigation of the Bidentate Pt-drugs DNA Binding --- p.129 / Chapter 4.5.5.1 --- Calculation Model --- p.129 / Chapter 4.5.5.2 --- Bidentate Pt-Drugs Reactions --- p.130 / Chapter 4.5.5.3 --- Selection of the Calculated Model for the QSAR Study --- p.133 / Chapter 4.5.5.4 --- QSAR Analysis Using ΔG+(guanine) for the Platinum Drugs with Bidentate Caboxylate Ligands --- p.136 / Chapter 4.5.5.5 --- QSAR Analysis Using ΔG(guanine) for the Platinum Drugs with Bidentate Carboxylate Ligands --- p.137 / Chapter 4.5.6 --- Conclusion --- p.138 / Chapter CHAPTER 5 --- Conclusion Remarks and Future Works / Chapter 5.1 --- Conclusion --- p.140 / Chapter 5.2 --- Future Works --- p.141 / REFERENCES --- p.142

慢性乙型肝炎病毒（HBV）感染是肝癌發生的重要因素，其中乙肝病毒X蛋白（HBx）在這一過程起著關鍵作用。研究發現，一些HBV變體和HBx突變具有更高致癌風險，而且這些變體和突變存在地區差異。香港是HBV感染高發地帶，因此本研究目的是從這一地區120個肝癌組織標本中篩查出HBx突變位點。我們用巢式PCR從84.16% (101/120)的標本中提取和擴增了HBx，並進行基因測序。三種HBx突變被檢測出，包括點突變，遠端羧基端截斷和缺失突變。其中點突變位點有39個，特別的是在50%的標本中檢測出A1630G/G1721A 和 A1762T/G1764A雙突變。在31.68% (32/101)的標本中發現遠端羧基端截斷，以及在2.97% (3/101)的標本中檢測出缺失突變。總之，大多數突變集中在HBx轉錄啟動域，表明這些突變在肝癌發生中可能起著重要作用。 / 缺氧誘導因數-1α（HIF-1α）在肝癌的發生和發展中也起著重要作用。研究發現，野生型HBx可以啟動HIF-1α，但是變異型HBx和HIF-1α的關係還沒有研究清楚。我們研究表明HBx轉錄啟動域是必需而且足夠啟動HIF-1α的。在這個區域的突變中，雙突變K130M/V131Z增強HBx對HIF-1α的活性，但遠端羧基端截斷和缺失突變削弱其功能。進一步研究發現，羧基端特別是119-140氨基酸對HBx的穩定和功能非常重要。肝癌標本中，我們也發現HBx和HIF-1α的表達呈正相關。因此，雖然不同的突變對於HBx的功能有不同的影響，但總的來說這些突變可以促進HIF-1α的表達和啟動，進而導致肝癌患者的預後不良。 / 靶向治療在肝癌綜合治療中扮演重要角色。索拉菲尼（Sorafenib）是一種多激酶抑制劑，臨床實驗發現它對晚期肝癌治療有效，但其抑制腫瘤血管生成機制還不完全清楚。我們研究發現Sorafenib明顯而且劑量依賴性地降低HIF-1α的表達和活化，進而抑制血管內皮生長因數（VEGF）的表達。Sorafenib抑制mTOR, ERK, p70S6K, RP-S6, eIF4E和4E-BP1等翻譯起始因數的磷酸化，從而抑制HIF-1α的合成而不影響其降解。體外實驗進一步發現Sorafenib降低HIF-1α和VEGF的表達，從而抑制腫瘤的血管形成和生長。總之，我們的研究表明sorafenib可能通過阻斷mTOR/p70S6K/4E-BP1 和 ERK 信號通路來抑制HIF-1α的合成，從而發揮其抗腫瘤血管生成作用。 / Chronic HBV infection is the leading cause of hepatocellular carcinoma (HCC) and HBx plays a crucial role in the molecular pathogenesis of HBV-related HCC. Previous investigations have indicated that some variations of HBV or mutations of HBx are associated with higher risk of HCC development, whereas the mutations profiles may be disparate in different regions. In the present studies, we thus aim to screen and identify the HBx mutation hotspots in 120 HCC tissues from Hong Kong, a region with HBV hyper-endemic. HBV DNAs were successfully isolated and amplified in 84.16% (101/120) HCC specimens via nest-PCR, and then subjected to gene sequencing. Three types of HBx mutations, including point mutations, distal carboxyl-terminal truncations and deletion mutations, were discovered. Among the point mutations, 39 mutation hotspots were indentified, with two double mutations (A1630G/G1721A and A1762T/G1764A) occurring in approximate 50% of 101 HCC cases. Distal C-terminal truncated mutations were discovered in 31.68% (32/101) of HCC cases, whereas deletion mutations were detected in 2.97% (3/101) of them. Overall, majority of identified mutations were located at the transactivation domain of HBx, suggesting the crucial roles of these mutations in HCC development. / Hypoxia-inducible factor-1α (HIF-1α) also closely involves in the development and progression of HCC. Wild-type HBx has been shown to activate HIF-1α. But the relationship between HBx mutants and activation of HIF-1α has not been fully elucidated. We here revealed that the transactivaiton domain of HBx was necessary and sufficient to activate HIF-1α. Double mutations K130M/V131Z in this domain enhanced the functionality of HBx in upregulating the expression and the activation of HIF-1α, whereas C-terminal truncations and deletion mutations weakened this prosperity of HBx. We further uncovered that the C-terminus, especially the region of amino acids 119-140, was essential for the stability and transactivation of HBx. The positive association between the HBx mutants and HIF-1α was found in the HCC tissue samples. Therefore, although mutations exerted different effects on the functionality of HBx, the overall activity of HBx mutants was suggested to upregulate HIF-1α, whose level is related to poor prognosis of HCC patients. / The therapy targeting a critical molecule in the development of HCC such as HIF-1α may be a potential and effective treatment regimen for HCC patients. Sorafenib, a multikinase inhibitor, has demonstrated promising results for the treatment of advanced HCC in clinical trials, but the mechanism that accounts for the anti-angiogenic efficiency of this agent has not been fully elucidated. We here revealed that sorafenib remarkably and dose-dependently decreased the expression and the transcriptional activity of HIF-1α, and its target gene, vascular endothelial grow factor (VEGF). Further analysis revealed that this reduction of HIF-1α by sorafenib was caused by the inhibition of HIF-1α protein synthesis rather than by the promotion of HIF-1α protein degradation. Moreover, the phosphorylated levels of mTOR, ERK, p70S6K, RP-S6, eIF4E and 4E-BP1 were significantly suppressed by sorafenib. In vivo studies further confirmed the inhibitory effect of sorafenib on the expression of HIF-1α and VEGF proteins, leading to a decrease of tumor vascularisation and growth. Collectively, our data suggest that sorafenib may exhibit anti-angiogenic activity by inhibiting HIF-1α synthesis, which is likely to be achieved through suppressing the phosphorylation of mTOR/p70S6K/4E-BP1 and ERK. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Liu, Liping. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 133-154). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract --- p.I / 摘要 --- p.IV / Publications --- p.VI / Acknowledgements --- p.VII / Abbreviations --- p.IX / List of Figures --- p.XI / List of Tables --- p.XIII / Table of Contents --- p.XIV / Chapter Chapter I --- General Introduction --- p.1 / Chapter 1.1 --- Overview of Hepatocellular Carcinoma --- p.1 / Chapter 1.2 --- HBV Infection and HCC Development --- p.6 / Chapter 1.3 --- Overview on Hepatitis B virus X Protein --- p.10 / Chapter 1.4 --- Roles of Hypoxia-inducible Factors in HCC --- p.17 / Chapter 1.5 --- Targeted Therapies and Sorafenib --- p.27 / Chapter Chapter II --- Identification of HBx Mutation Hotspots in HCC Tissues --- p.31 / Chapter 2.1 --- Abstract --- p.31 / Chapter 2.2 --- Introduction --- p.32 / Chapter 2.3 --- Materials and Methods --- p.35 / Chapter 2.4 --- Results --- p.40 / Chapter 2.5 --- Discussion --- p.53 / Chapter Chapter III --- The Relationship between HBx Mutants and HIF-1α --- p.59 / Chapter 3.1 --- Abstract --- p.59 / Chapter 3.2 --- Introduction --- p.60 / Chapter 3.3 --- Materials and Methods --- p.63 / Chapter 3.4 --- Results --- p.70 / Chapter 3.5 --- Discussion --- p.91 / Chapter Chapter IV --- The Effects of Sorafenib on Hypoxia-inducible Factor-1α --- p.96 / Chapter 4.1 --- Abstract --- p.96 / Chapter 4.2 --- Introduction --- p.98 / Chapter 4.3 --- Materials and Methods --- p.101 / Chapter 4.4 --- Results --- p.108 / Chapter 4.5 --- Discussion --- p.124 / Chapter Chapter V --- Conclusion and Future Plans --- p.129 / Chapter 5.1 --- Conclusion --- p.129 / Chapter 5.2 --- Future Plans --- p.131 / References --- p.133

Both mRNA and protein expression levels of P-glycoprotein, one of the major factors involved in drug resistance, was decreased in Pa-treated R-HepG2 cells. The chemo-sensitivity of these MDR cells towards doxorubicin would be enhanced by pretreatment of Pa. / In the study, 35 TCMs with historical background in treating liver diseases were screened. S. barbata was chosen for intensive studies based on its significant anti-hepatoma activity. Using bioassay-guided purification approach, an active component, pheophorbide a (Pa) - a chlorophyll derivative, was isolated from Scutellaria barbata. / Motivated by the severe health hazards worldwide caused by liver cancer, and the pronounced side effects of some recent anti-hepatoma agents in clinical treatment, we have initiated a research project in screening safe and effective agents from Traditional Chinese Medicine (TCM) for the treatment of hepatoma. The main objective of this research is to define the in vitro and in vivo anti-proliferative activities and to identify the action mechanisms of a TCM, the aerial part of Scutellaria barbata , in human hepatoma cells (HepG2 and Hep3B cells). / Pa exhibited anti-proliferative effects on HepG2 and Hep3B cells, through cell-cycle arrest and apoptosis, with IC50 values being 12.5 and 25.7 muM respectively. However, Pa produced insignificant cytotoxic effect on WRL-68 cells, a normal hepatic cell line. Pa also caused cell death in R-HepG2 cells, a multi-drug resistant (MDR) cell line developed from HepG2 cells. Microarray analysis indicated that a hypothetical protein FLJ10803 was found to be down-regulated upon the treatment of Pa on HepG2 cells. The sub-cellular localization of FLJ10803 was demonstrated by over-expression of the GFP fusion protein in HepG2 cells. / The anti-tumor effects of Pa could be enhanced by photodynamic therapy (PDT) approach, presumably due to the rapid generation of reactive oxygen species in the drug-binding site. Pa-PDT showed potent cytotoxicity on hepatoma cell lines, HepG2 and Hep3B, with IC50 values being 0.4 and 1.5 muM, respectively. The antitumor effects were confirmed by studies using animal model, where Pa treatment (300mug/kg/day, s.c.) could significantly inhibit the growth of Hep3B cells in nude mice after PDT treatment in vivo. Fluorescent imaging showed that Pa was located at the mitochondria, and the induction of cell death was found to be initiated by the mitochondrial dependent apoptotic pathway. Results of 2D-gel analysis suggested that Pa-PDT activated an immune-marker expression pathway that results in an over expression of HLA class I proteinsin Pa-PDT treated HepG2 cells. / To conclude, Pa may be a candidate for further development into an anti-hepatomic agent for clinical application. / Tang, Ming Kuen. / "September 2007." / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4742. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 227-243). / 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. / Abstracts in English and Chinese. / School code: 1307.

Fung Yin Lee, Annie. / "January 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 132-146). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2009. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 4 uppsatser.

published_or_final_version / abstract / Chemistry / Doctoral / Doctor of Philosophy