Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2006. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Dissertation (Ph.D.)--University of Toledo, 2008. / "In partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biomedical Sciences." Title from title page of PDF document. Bibliography: pages 73-77, 93-95, 109-111, 145-172.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2010.

No description available.

By using the acridine orange (AO) staining method to examine the release of contents from lysosomes, it was found that PD released AO into the cytosol in both cell lines. However, the releasing pattern of HepG2 and RHepG2 was quite different. Upon PD treatment, the release of AO in HepG2 cells was graduate and slow while that in RHepG2 was sudden and sharp. / Cancer is one of the leading causes of death in the world. During cancer treatment, development of multidrug resistance (MDR) is always the major cause of failures of chemotherapy in human cancers. In our project, hepatocarcinoma HepG2 and its drug-resistant derivatives RHepG2 with MDR towards doxorubicin (Dox), fenretinide and Taxol were used to examine the differences in their response towards various anti-cancer agents. / From the AO staining, most of the lysosomes were found in the cytosol near the nucleus. However, some lysosomes were found inside the nucleus occasionally. When we double stained the HepG2 cells with DiOC6(3), it was found that the lysosomes were actually located inside the nuclear tubules. However, no such lysosome migration was observed after treating the HepG2 cells with PD. Thus, lysosomes inside the nuclear tubules might not be involved in the PD-induced lysosomal pathway. The mechanism that leads to the migration of lysosomes into the nuclear tubules is still unclear. / From the Western blot analysis, cathepsin D (Cat D) and cathepsin L (Cat L) were both released from the lysosomes after treating the two cell lines with PD. Also, it seemed likely that Cat L was released earlier than that of cyt c. This implies that lysosomal permeabilization is an early event in apoptosis. With the use of siRNA technology, it was found that RHepG2 with the knockdown of Cat D and Cat L were more tolerant and vulnerable towards PD, respectively. These suggest that Cat D and Cat L might act oppositely in the apoptotic pathway. Furthermore, the addition of Cat D inhibitor, pepstatin A, blocked the PD-mediated cell death in RHepG2 cells further confirms that Cat D is a pro-apoptotic protein that is involved in the apoptotic pathway. / In conclusion, PD was a potent anti-cancer agent that could reverse the MDR properties of RHepG2 and kill more RHepG2 cells through lysosomal and mitochondrial apoptotic pathway. / Next, we investigated the underlying killing mechanism and found out that PD switched on both the mitochondrial and lysosomal apoptotic pathway in both cell lines. Our results indicate that PD was able to depolarize mitochondrial membrane potential and release apoptosis inducing factor (AIF) and cytochrome c (cyt c) from the mitochondria to cytosol. Also, PD was able to act on isolated mitochondria directly, causing a stronger mitochondrial membrane permeabilization and more AIF release from the RHepG2 than that of the parental cells. / Polyphyllin D (PD) is a saponin found in a tradition Chinese herb, Paris polyphylla, which has been used to treat liver cancers in China for many years. Interestingly, from the MTT assays, we found out that RHepG2 (IC50: 2.0 muM) was more sensitive towards PD when compared to that of its parental cells (IC50: 3.9 muM). To keep the MDR properties, RHepG2 cells were routinely cultured with 1.2 muM of Dox. When we cultured RHepG2 in the absence of Dox but with 1.2 muM of PD for 28 days, the Pgp expression could not be maintained. However, such high expression level of Pgp was maintained when RHepG2 cells were treated with vincristine (1.2 muM) in the absence of Dox. This indicates that vincristine was a substrate of Pgp to keep the Pgp expression in RHepG2 cells while PD was not. / When incubated with different concentrations of Dox, RHepG2 accumulated less Dox than that of its parental HepG2 cells. When probed by the antibody against P-glycoprotein (Pgp), RHepG2 showed a strong Pgp expression. With the addition of Pgp modulator, verapamil, RHepG2 accumulated more Dox. All these findings indicate that Pgp is a mediator giving rise the MDR in RHepG2 cells. However, RHepG2 had a higher resistance to Dox than its parental line even co-cultured with verapamil. RHepG2 remained viable at the intracellular Dox concentration that was toxic to HepG2 cells. These observations suggest that the MDR properties of RHepG2 involved multiple mechanisms in addition to the effect of Pgp. / Lee, Kit Ying Rebecca. / "August 2007." / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4735. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 241-253). / 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.

Li Ka Wai Kay. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references. / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT (ENGLISH/CHINESE) --- p.ii / TABLE OF CONTENTS --- p.viii / LIST OF FIGURES --- p.xii / LIST OF APPENDICES --- p.xiii / Chapter 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Gastric carcinomas --- p.1 / Chapter 1.1a --- Epidemiology --- p.1 / Chapter 1.1b --- Classification --- p.2 / Chapter 1.1c --- TNM staging --- p.3 / Chapter 1.1d --- Prognosis --- p.4 / Chapter 1.1e --- Etiology --- p.6 / Chapter 1.1f --- Genetic alteration in gastric cancer --- p.10 / Chapter 1.2 --- Treatment --- p.16 / Chapter 1.2a --- "Surgery, chemotherapy, and others" --- p.16 / Chapter 1.2b --- Response rate of treatments in previous studies --- p.18 / Chapter 1.2c --- Chemotherapeutic Drugs --- p.21 / Chapter 1.2c (1) --- 5-fluorouracil (5-FU) --- p.22 / Chapter 1.2c (2) --- cis-diamminedichloroplatinum (Cisplatin) --- p.23 / Chapter 1.2c (3) --- Doxorubicin (Adriamycin) --- p.23 / Chapter 1.2c (4) --- Daunorubicin --- p.25 / Chapter 1.2c (5) --- Epirubicin --- p.25 / Chapter 1.2d --- Toxicity of chemotherapeutic drugs --- p.26 / Chapter 1.2d (1) --- Side effects of 5-FU --- p.26 / Chapter 1.2d (2) --- "Side effects of anthracyc lines (adriamycin, daunobicin, epuirbicin)" --- p.27 / Chapter 1.2d (3) --- Side effects of cisplatin --- p.28 / Chapter 1.3 --- Mechanisms of drug resistance --- p.28 / Chapter 1.3a --- Drug resistance --- p.28 / Chapter 1.3b --- P-glycoprotein (MDR1 gene) --- p.29 / Chapter 1.3c --- p53 tumor suppressor gene --- p.35 / Chapter 1.4 --- Chemosensitivity testing --- p.40 / Chapter 1.4a --- Original of chemosensitivity testing --- p.40 / Chapter 1.4b --- Non-clonogentic assay --- p.40 / Chapter 1.4c --- Clonogenic assay --- p.42 / Chapter 2 --- AIM OF MY STUDY --- p.44 / Chapter 3 --- MATERIALS AND METHODS --- p.45 / Chapter 3.1 --- Patients --- p.45 / Chapter 3.2 --- Tumor collection and handling procedure --- p.46 / Chapter 3.2a --- Large tumor tissue from gastrectomy --- p.46 / Chapter 3.2b --- Pseudo-biopsies --- p.47 / Chapter 3.3 --- Chemosensitivity testing --- p.48 / Chapter 3.3a --- Cell Plating --- p.48 / Chapter 3.3b --- Drug testing --- p.49 / Chapter 3.4 --- Chemosensitivity analysis --- p.50 / Chapter 3.5 --- Conformational sensitive gel electrophoresis analysis (CSGE) and single strand conformational polymorphism (SSCP) --- p.51 / Chapter 3.5a --- Preparation of genomic DNA --- p.51 / Chapter 3.5b --- PCR condition for CSGE analysis --- p.51 / Chapter 3.5c --- Scanning PCR products by CSGE --- p.52 / Chapter 3.5d --- PCR condition for SSCP analysis --- p.53 / Chapter 3.5e --- Scanning PCR products by SSCP --- p.53 / Chapter 3.6 --- Reverse transcription-polymerase chain reaction (RT-PCR) for multi-drug drug resistance (MDR1) gene --- p.54 / Chapter 3.6a --- Isolation of RNA --- p.54 / Chapter 3.6b --- cDNA synthesis --- p.55 / Chapter 3.6c --- PCR primers --- p.55 / Chapter 3.6d --- Optimalization of PCR condition for MDR1 gene expression --- p.56 / Chapter 3.6e --- PCR of β2-m gene --- p.57 / Chapter 3.6f --- PCR of MDR1 gene and analysis of its expression --- p.57 / Chapter 3.7 --- Immunohistochemistry --- p.58 / Chapter 3.7a --- Immunostaining by DO-7 --- p.58 / Chapter 3.7b --- lmmunohistochemistochemical analysis of p53 protein expression --- p.59 / Chapter 3.8 --- Statistics --- p.59 / Chapter 4. --- RESULTS --- p.60 / Chapter 4.1 --- Chemosensitivity testing --- p.60 / Chapter 4.1a --- Tests completed --- p.60 / Chapter 4.1b --- Number of cases tested for each drug --- p.60 / Chapter 4.1c --- 〇D reading of the background samples --- p.60 / Chapter 4.1d --- Dose-dependent response curve --- p.61 / Chapter 4.1e --- Unique IC50 for each tumor in each drug test --- p.61 / Chapter 4.1f --- Wide distribution of ic50 for anti-tumor drugs --- p.61 / Chapter 4.1g --- Chemosensitivity and tumor histologic type --- p.63 / Chapter 4.1h --- Correlation of ic50 with tumor stage --- p.63 / Chapter 4.2 --- Immunohistochemical staining of p53 protein (DO-7) --- p.64 / Chapter 4.2a --- p53 protein accumulation in samples --- p.64 / Chapter 4.2b --- Correlation of p53 IHC expression and chemosensitivity --- p.64 / Chapter 4.3 --- SSCP and CSGE --- p.65 / Chapter 4.3a --- Detection of abnormal band movement --- p.65 / Chapter 4.3b --- Correlation of p53 mutations with chemosensitivity --- p.66 / Chapter 4.3c --- Concordance between IHC and SSCP/CSGE --- p.66 / Chapter 4.4 --- MDR1 gene expression --- p.67 / Chapter 4.4a --- MDR1 gene expression in normal and tumors --- p.67 / Chapter 4.4b --- Correlation of MDR1 expression and chemosensitivity --- p.68 / Chapter 4.5 --- Pseudobiopsies --- p.68 / Chapter 5 --- DISCUSSION --- p.70 / Chapter 5.1 --- p53 analysis of the tumors --- p.70 / Chapter 5.1a --- Immunohistochemistry versus mutational analysis --- p.70 / Chapter 5.1b --- Methods of mutational analysis --- p.73 / Chapter 5.1c --- Comparing IHC results with previous findings --- p.77 / Chapter 5.1d --- Comparing SSCP/ CSGE results with previous findings --- p.78 / Chapter 5.1e --- Correlation of IHC and SSCP/CSGE results --- p.81 / Chapter 5.2 --- MDR1 expression --- p.85 / Chapter 5.2a --- Methods for detecting MDR1 expression --- p.85 / Chapter 5.2b --- Comparing MDR1 expression results with published data --- p.88 / Chapter 5.2c --- Correlation between chemosensitivity and MDR1 gene expression --- p.92 / Chapter 5.3 --- Chemosensitivity testing --- p.94 / Chapter 5.3a --- Chemosensitivity testing method --- p.94 / Chapter 5.3b --- The chemosensitivity results --- p.102 / Chapter 5.3c --- Chemosensitivity and MDR1 expression --- p.108 / Chapter 5.3d --- Chemosensitivity and p53 immunohistochemical expression… --- p.110 / Chapter 5.3e --- Chemosensitivity and p53 mutations --- p.112 / Chapter 5.3f --- Limitation of this study --- p.115 / Chapter 5.3g --- Pseudobiopsies and large tumor samples --- p.118 / Chapter 6. --- conclusions --- p.121 / figures / appendices / references

Cheung, Ka Yan. / "Aug 2010." / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 157-167). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgments --- p.v / Table of Contents --- p.vi / List of Figures --- p.x / List of Tables --- p.xi / Abbreviations --- p.xii / Chapter Chapter1 --- General Introduction --- p.1 / Chapter 1.1 --- Cancer epidemiology and managements --- p.2 / Chapter 1.2 --- Photodynamic therapy (PDT) as cancer treatment --- p.7 / Chapter 1.3 --- Pheophorbide a (Pa) as a photosensitizer for PDT --- p.13 / Chapter 1.4 --- Aim of study --- p.15 / Chapter Chapter2 --- The anti-proliferative effect of pheophorbide a- mediated photodynamic therapy on human breast adenocarcinoma cell line MCF-7 --- p.17 / Chapter 2.1 --- Introduction / Chapter 2.1.1 --- Cell cycle regulation --- p.18 / Chapter 2.1.2 --- Growth arrest and DNA damage inducible (GADD) genes as cell cycle regulators --- p.22 / Chapter 2.2 --- Materials and Methods / Chapter 2.2.1 --- Materials / Chapter 2.2.1.1 --- Cell line --- p.29 / Chapter 2.2.1.2 --- "Cell culture medium, supplements and other reagents" --- p.29 / Chapter 2.2.1.3 --- Gene expression assay reagents --- p.30 / Chapter 2.2.1.4 --- Reagents and buffers for Western blotting --- p.32 / Chapter 2.2.1.5 --- Cell cycle analysis reagents --- p.35 / Chapter 2.2.2 --- Methods / Chapter 2.2.2.1 --- Cell line propagation and subculture --- p.36 / Chapter 2.2.2.2 --- Whole-transcript expression micro array analysis --- p.37 / Chapter 2.2.2.3 --- GADD genes expression assay- RT-PCR --- p.37 / Chapter 2.2.2.4 --- Cell cycle analysis --- p.40 / Chapter 2.2.2.5 --- Western Blotting --- p.41 / Chapter 2.2.2.6 --- Statistical analysis --- p.43 / Chapter 2.3 --- Results / Chapter 2.3.1 --- Effect of Pa-PDT on GADD genes expression by whole-transcript expression microarray analysis --- p.44 / Chapter 2.3.2 --- Effect of Pa-PDT on GADD genes expression by RT-PCR --- p.46 / Chapter 2.3.3 --- Temporal change in the cell cycle profile after Pa-PDT --- p.48 / Chapter 2.3.4 --- Effect of Pa-PDT on cell cycle associated proteins --- p.65 / Chapter 2.4 --- Discussion --- p.67 / Chapter Chapter3 --- Development of drug resistance in human breast adenocarcinoma cell line MDA and the circumvention by pheophorbide a-mediated photodynamic therapy --- p.77 / Chapter 3.1 --- Introduction / Chapter 3.1.1 --- Clinical Importance of multidrug resistance (MDR) --- p.78 / Chapter 3.1.2 --- Mechanisms of MDR --- p.78 / Chapter 3.1.3 --- Development of MDR cell lines --- p.82 / Chapter 3.1.4 --- Reversal of MDR by P-glycoprotein modulators --- p.83 / Chapter 3.1.5 --- Therapeutic potential of Pa-PDT in treating MDR cancers --- p.83 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Materials / Chapter 3.2.1.1 --- Cell line --- p.85 / Chapter 3.2.1.2 --- "Cell culture medium, supplements and other reagents" --- p.85 / Chapter 3.2.1.3 --- Cell viability assay reagents --- p.85 / Chapter 3.2.1.4 --- Gene expression assay reagents --- p.86 / Chapter 3.2.2 --- Methods / Chapter 3.2.2.1 --- Cell line propagation and subculture --- p.87 / Chapter 3.2.2.2 --- Drug-resistance development --- p.88 / Chapter 3.2.2.3 --- Measurement of cell viability - MTT reduction assay --- p.88 / Chapter 3.2.2.4 --- ABCB1 expression assay- RT-PCR --- p.89 / Chapter 3.2.2.5 --- Doxorubicin uptake assay --- p.91 / Chapter 3.2.2.6 --- Pheophorbide a uptake assay --- p.91 / Chapter 3.2.2.7 --- Statistical analysis --- p.92 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Cytotoxicity of doxorubicin on MDA and MDA-R cells --- p.93 / Chapter 3.3.2 --- mRNA expression of ABCB1 (P-glycoprotein) in MDA and MDA-R cells --- p.96 / Chapter 3.3.3 --- Doxorubicin uptake by MDA and MDA-R cells --- p.98 / Chapter 3.3.4 --- Circumvention of drug resistance in MDA-R cells by Pa-PDT --- p.102 / Chapter 3.3.5 --- Pheophorbide a uptake by MDA and MDA-R cells --- p.104 / Chapter 3.4 --- Discussion --- p.106 / Chapter Chapter4 --- Synergistic anti-proliferation of pheophorbide a-mediated photodynamic therapy and doxorubicin on multidrug resistant uterine sarcoma cell line Dx5 --- p.113 / Chapter 4.1 --- Introduction / Chapter 4.1.1 --- Clinical limitations of doxorubicin as chemotherapeutic drug --- p.114 / Chapter 4.1.2 --- Clinical limitations of photodynamic therapy --- p.115 / Chapter 4.1.3 --- Combination therapy with Dox and Pa-PDT --- p.117 / Chapter 4.1.4 --- Uterine sarcoma cell line Dx5 as in vitro model for combination therapy --- p.118 / Chapter 4.2 --- Materials and Methods / Chapter 4.2.1 --- Materials / Chapter 4.2.1.1 --- Cell line --- p.120 / Chapter 4.2.1.2 --- "Cell culture medium, supplements and other reagents" --- p.120 / Chapter 4.2.1.3 --- Anti-cancer drugs --- p.121 / Chapter 4.2.1.4 --- "ROS inhibitor, α-tocopherol" --- p.121 / Chapter 4.2.1.5 --- Cell viability assay reagents --- p.122 / Chapter 4.2.1.6 --- P-glycoprotein activity assay reagents --- p.122 / Chapter 4.2.2 --- Methods - / Chapter 4.2.2.1 --- Cell line propagation and subculture --- p.123 / Chapter 4.2.2.2 --- Cell viability assay --- p.123 / Chapter 4.2.2.3 --- P-glycoprotein activity assay --- p.124 / Chapter 4.2.2.4 --- Statistical analysis --- p.125 / Chapter 4.3 --- Results / Chapter 4.3.1 --- Combination therapy of Pa-PDT and doxorubicin in Dx5 cells --- p.126 / Chapter 4.3.2 --- Effect of α-tocopherol on the synergism between Pa-PDT and doxorubicin in Dx5 cells --- p.129 / Chapter 4.3.3 --- Effect of Pa-PDT on P-glycoprotein activity in Dx5 cells --- p.132 / Chapter 4.3.4 --- Combination therapy of Pa-PDT and doxorubicin in SA cells --- p.138 / Chapter 4.4 --- Discussion --- p.141 / Chapter Chapter5 --- General Discussion --- p.148 / Chapter 5.1 --- Pa-PDT induced growth arrest and DNA fragmentation in breast cancer MCF-7 cells --- p.149 / Chapter 5.2 --- Circumvention of doxorubicin resistance by Pa-PDT in breast cancer MDA cells --- p.151 / Chapter 5.3 --- Synergistic anti-proliferation of Pa-PDT and doxorubicin on uterine sarcoma cell line Dx5 --- p.151 / Chapter 5.4 --- Clinical implication --- p.153 / Chapter 5.5 --- Conclusions and future perspectives --- p.153 / References --- p.157