Chow Chit. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 130-156). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Table of Contents --- p.vii / List of Tables --- p.xi / List of Figures --- p.xii / List of Abbreviations --- p.xiv / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Malignant Lymphoma --- p.1 / Chapter 1.2 --- Non-Hodgkin's Lymphoma --- p.1 / Chapter 1.3 --- NK/T Cell Lymphoma --- p.2 / Chapter 1.3.1 --- General Features of NK/T Cell Lymphoma --- p.2 / Chapter 1.3.2 --- Histology of NK/T Cell Lymphoma --- p.3 / Chapter 1.3.3 --- Subtypes NK/T Cell Lymphoma --- p.4 / Chapter 1.3.4 --- Overview of NK/T Cell Lymphoma Cell Lines --- p.5 / Chapter 1.3.4.1 --- NK/T Cell Lymphoma Cell Lines-NK-92 and SNK-6 --- p.6 / Chapter 1.3.5 --- NK/T Cell Lymphoma and Interleukins --- p.8 / Chapter 1.4 --- Interleukin-2 and Interleukin-15 --- p.9 / Chapter 1.5 --- IL-2 and IL-15 Receptor --- p.10 / Chapter 1.6 --- Cellular Signaling Pathways Regulated by IL-2 and IL-15 --- p.11 / Chapter 1.6.1 --- JAK/STAT Pathway --- p.12 / Chapter 1.6.2 --- PI3K/Akt Pathway --- p.14 / Chapter 1.7 --- Epstein-Barr Virus: an Oncogenic Virus --- p.20 / Chapter 1.7.1 --- Overview of EBV --- p.20 / Chapter 1.7.2 --- Epidemiology --- p.20 / Chapter 1.7.3 --- Life Cycle of EBV --- p.21 / Chapter 1.7.4 --- Latency Infection of EBV --- p.21 / Chapter 1.7.5 --- Role of EBV latent genes in oncogenesis --- p.23 / Chapter 1.7.5.1 --- EBER1 and 2 --- p.23 / Chapter 1.7.5.2 --- EBNAs --- p.24 / Chapter 1.7.5.3 --- LMPs --- p.25 / Chapter 1.7.6 --- Lytic Cycle of EBV --- p.26 / Chapter 1.7.7 --- Signaling Pathways and EBV --- p.27 / Chapter Chapter 2: --- Aim of Study --- p.29 / Chapter Chapter 3: --- Materials and Methods --- p.31 / Chapter 3.1 --- IL-2 and IL-15 on NK/T Cell Lymphoma Cell Lines and patients --- p.31 / Chapter 3.1.1 --- Cell Lines Maintenance --- p.31 / Chapter 3.1.2 --- Patients --- p.32 / Chapter 3.2 --- "Assays of IL-2, IL-15 and IFN-γ in culture supernatants and patient sera" --- p.32 / Chapter 3.2.1 --- IL-2 ELISA --- p.32 / Chapter 3.2.2 --- IL-15 ELISA --- p.33 / Chapter 3.2.3 --- IFN-γ ELISA --- p.34 / Chapter 3.3 --- Effect of IL-2 and IL-15 on NK/T Cell Lymphoma Cell Lines --- p.35 / Chapter 3.3.1 --- Cell Growth and Viability Determination --- p.35 / Chapter 3.3.2 --- Apoptosis Assays on Interleukin-starved NK-92 cells --- p.35 / Chapter 3.3.2.1 --- DNA Laddering Analysis --- p.36 / Chapter 3.3.2.2 --- Cell Cycle and Apoptosis Determination by PI Staining --- p.37 / Chapter 3.3.2.3 --- Caspase 3 Activity Assay --- p.37 / Chapter 3.4 --- PI3K/Akt Pathway Study --- p.39 / Chapter 3.4.1 --- Determination of AKT1 Gene Amplification by Real-Time Quantitative PCR --- p.39 / Chapter 3.4.1.1 --- DNA Extraction for Real-Time Quantitative PCR --- p.39 / Chapter 3.4.1.2 --- AKT1 Real-Time Quantitative PCR --- p.40 / Chapter 3.4.2 --- Determination of Akt Expression --- p.41 / Chapter 3.4.2.1 --- Normal NK Cell Purification from Buffy Coat --- p.41 / Chapter 3.4.2.2 --- Determination of the Purity of Extracted NK Cells --- p.43 / Chapter 3.4.2.3 --- Interleukin Treatment of Normal NK Cells and NK-92 --- p.43 / Chapter 3.4.2.4 --- Protein Extraction and Western Blot Analysis --- p.43 / Chapter 3.4.3 --- Study of PI3 K/Akt pathway using PI3K inhibitor --- p.47 / Chapter 3.4.3.1 --- Cell Growth and Viability Assay --- p.47 / Chapter 3.4.3.2 --- Apoptosis Assay by DNA Laddering and Pi-Staining on NK-92 cells --- p.48 / Chapter 3.4.3.3 --- Determination of activated Akt after LY294002 and Wortmannin treatment --- p.48 / Chapter 3.5 --- Effect of IL-2 and IL-15 on the JAK/STAT pathway and PDK/Akt pathway of NK/T Cell Lymphoma Cell Lines --- p.49 / Chapter 3.5.1 --- Cell Treatment --- p.49 / Chapter 3.5.2 --- Study of JAK/STAT and PI3K/Akt Pathways by Western Blotting --- p.49 / Chapter 3.5.3 --- "Assays of IL-2, IL-15 and IFN-γ in the NK-92 Cell Culture Medium by ELISA" --- p.50 / Chapter 3.5.4 --- Determination of EBV Status after IL-2 and IL-15 Treatment --- p.51 / Chapter 3.5.4.1 --- RNA Extraction --- p.51 / Chapter 3.5.4.2 --- Reverse-transcriptase Reaction --- p.52 / Chapter 3.5.4.3 --- PCR for EBV-related Genes --- p.53 / Chapter 3.5.4.4 --- EBER-ISH --- p.54 / Chapter 3.5.4.5 --- Real-time Quantitative PCR for EBER1 --- p.56 / Chapter 3.5.4.6 --- Western Blot for LMP1 --- p.56 / Chapter 3.6 --- Statistical Analysis --- p.57 / Chapter Chapter 4: --- Results --- p.59 / Chapter 4.1.1 --- "IL-2, IL-15 and IFN-γ Levels in the Serum of Patients with NK/T Cell Lymphoma" --- p.59 / Chapter 4.1.2 --- IL-2 and IL-15 Level in Culture Supernatant of NK-92 --- p.59 / Chapter 4.1.3 --- IFN-γ induction in supernatant of NK-92 --- p.60 / Chapter 4.2 --- Effect of IL-2 and IL-15 on NK/T Cell Lymphoma Cell Lines --- p.61 / Chapter 4.2.1 --- Cell Growth and Viability --- p.61 / Chapter 4.2.2 --- Apoptosis Study of Interleukin-starved NK-92 --- p.62 / Chapter 4.2.2.1 --- DNA fragmentation and Cell Cycle studies --- p.62 / Chapter 4.2.2.2 --- Caspase 3 Activity in NK-92 --- p.62 / Chapter 4.3 --- Akt in NK-92 --- p.63 / Chapter 4.3.1 --- Confirmation ofAKTl Amplification in NK-92 --- p.63 / Chapter 4.3.2 --- Akt Protein Quantification in NK-92 cells --- p.63 / Chapter 4.3.3 --- Activated Akt and STAT proteins in IL-2 or IL-15 stimulated NK-92 and normal NK cells --- p.64 / Chapter 4.4. --- PI3K/Akt Pathway --- p.64 / Chapter 4.4.1 --- Phosphorylation of Components of the PI3K/Akt Pathway --- p.64 / Chapter 4.4.2. --- Role of PI3K/Akt Pathway in NK/T Cell Lymphoma Cell Lines --- p.65 / Chapter 4.4.2.1 --- Cell Growth and Viability Studies --- p.65 / Chapter 4.4.2.2 --- Apoptosis and Cell Cycle Arrest Induction by LY294002 in NK-92 --- p.66 / Chapter 4.4.2.3 --- Confirmation of the effect of LY294002 on the PDK/Akt pathway in NK-92 cells --- p.67 / Chapter 4.5 --- Phosphorylation of STAT family proteins --- p.67 / Chapter 4.6 --- Regulation of EBV-related genes in NK-92 --- p.68 / Chapter Chapter 5: --- Discussion --- p.71 / Chapter 5.1 --- Cytokine level in patient serum --- p.71 / Chapter 5.2 --- Source of IL-2 and IL-15 for NK-92 cells --- p.72 / Chapter 5.3 --- Induction of IFN-γ in NK-92 --- p.73 / Chapter 5.4 --- Role of IL-2 and IL-15 on NK/T cell lymphoma cell lines --- p.75 / Chapter 5.4.1 --- Cell Growth and Viability Maintenance by IL-2 and IL-15 --- p.75 / Chapter 5.4.2 --- Apoptosis induced by interleukin-starving in NK-92 --- p.75 / Chapter 5.5 --- Aberrant activation of signaling pathways by IL-2 or IL-15 in NK-92 --- p.77 / Chapter 5.5.1 --- Hypersensitivity of NK-92 cells to IL-2 or IL-15 --- p.77 / Chapter 5.5.2 --- PI3K/Akt pathway in NK/T cell lymphoma cell lines --- p.78 / Chapter 5.5.2.1 --- Confirmation of AKT1 amplification --- p.78 / Chapter 5.5.2.2 --- Constitutive activation of Akt in NK/T cell lymphoma cell lines --- p.79 / Chapter 5.5.2.3 --- Role of PI3K/Akt in NK/T cell lymphoma cell lines --- p.80 / Chapter 5.5.2.4 --- IL-2 and IL-15 induce differential sensitivity of NK-92 to LY294002 --- p.81 / Chapter 5.5.2.5 --- NK/T cell lymphoma cell lines are wortmannin-insensitive --- p.82 / Chapter 5.6 --- Jak/STAT pathway in NK/T cell lymphoma cell lines --- p.83 / Chapter 5.6.1 --- STAT3 and STAT5 were activated by both IL-2 and IL-15 --- p.83 / Chapter 5.6.2 --- STAT6 was activated in NK/T cell lymphoma cell lines --- p.84 / Chapter 5.6.3 --- "Differential regulation of STAT 1, STAT3 (Ser-727) and STAT6 in NK/T cell lymphoma cell lines" --- p.85 / Chapter 5.6 --- EBV gene regulation in NK-92 --- p.87 / Chapter Chapter 6: --- Conclusion --- p.91 / Tables --- p.93 / Figures --- p.102 / Reference --- p.128