Resistance to drug-induced apoptosis in T-cell acute lymphoblastic leukemia.

Leung Kam Tong.Thesis submitted in: September 2006.Thesis (M.Phil.)--Chinese University of Hong Kong, 2007.Includes bibliographical references (leaves 79-95).Abstracts in English and Chinese.Abstract --- p.iAbstract (Chinese) --- p.iiiAcknowledgements --- p.vTable of contents --- p.viList of figures --- p.ixList of abbreviations --- p.xiiChapter Chapter 1 --- General Introduction --- p.1Chapter 1.1 --- Acute lymphoblastic leukemia --- p.1Chapter 1.2 --- T-cell acute lymphoblastic leukemia --- p.2Chapter 1.2.1 --- Chemotherapy --- p.2Chapter 1.2.1.1 --- Induction therapy --- p.2Chapter 1.2.1.2 --- Intensification therapy --- p.3Chapter 1.2.1.3 --- Maintenance therapy --- p.3Chapter 1.2.2 --- Chemoresistance in T-ALL --- p.3Chapter 1.3 --- Apoptosis and chemoresistance --- p.5Chapter 1.3.1 --- "Initiation, execution and regulation of apoptosis" --- p.5Chapter 1.3.1.1 --- Initiation of apoptosis --- p.5Chapter 1.3.1.2 --- Execution of apoptosis --- p.7Chapter 1.3.1.3 --- Regulation of apoptosis --- p.7Chapter 1.3.2 --- Mechanisms of resistance to apoptosis --- p.9Chapter 1.3.2.1 --- Overexpression of pro-survival proteins --- p.9Chapter 1.3.2.2 --- Downregulation and mutation of pro-apoptotic proteins --- p.11Chapter 1.3.2.3 --- Other mechanisms --- p.13Chapter 1.4 --- Bcl-2 interating mediator of cell death --- p.14Chapter 1.4.1 --- Role of Bim in apoptosis --- p.16Chapter 1.4.2 --- Regulation of Bim --- p.17Chapter 1.4.2.1 --- Transcriptional regulation of Bim --- p.18Chapter 1.4.2.2 --- Post-transcriptional regulation of Bim --- p.18Chapter 1.5 --- c-Jun N-terminal kinase --- p.20Chapter 1.5.1 --- Pro-apoptotic role of JNK --- p.21Chapter 1.5.2 --- Anti-apoptotic role of JNK --- p.21Chapter 1.6 --- Hypotheses --- p.22Chapter Chapter 2 --- Materials and Methods --- p.23Chapter 2.1 --- Cell culture --- p.23Chapter 2.2 --- Induction of quantification of apoptosis --- p.24Chapter 2.3 --- Determination of caspase activities --- p.24Chapter 2.4 --- Western blotting --- p.25Chapter 2.4.1 --- Protein extraction and determination of protein concentration --- p.25Chapter 2.4.2 --- SDS-PAGE and immunodetection --- p.26Chapter 2.5 --- Cell-free apoptosis reactions --- p.27Chapter 2.6 --- Analysis of mitochondrial membrane potential --- p.27Chapter 2.7 --- Transient transfection of Sup-Tl cells --- p.28Chapter 2.8 --- Reverse transcription-polymerase chain reaction (RT-PCR) --- p.28Chapter 2.8.1 --- RNA isolation --- p.28Chapter 2.8.2 --- Synthesis of first-strand cDNA --- p.29Chapter 2.8.3 --- Polymerase chain reaction --- p.29Chapter 2.9 --- Alkaline phosphatase digestion of Bim --- p.30Chapter Chapter 3 --- Results --- p.31Chapter 3.1 --- The T-ALL cell line Sup-Tl is resistant to etoposide-induced apoptosis --- p.31Chapter 3.2 --- Sup-Tl cells are resistant to etoposide-induced caspase activation --- p.40Chapter 3.3 --- Sup-Tl cells are insusceptible to etoposide-induced mitochondrial alterations --- p.46Chapter 3.4 --- BimEL is required for etoposide-induced apoptosis in Sup-Tl cells --- p.51Chapter 3.5 --- The reduced level of BimEL in Sup-Tl cells is owing to the presence of constitutively active JNK --- p.58Chapter Chapter 4 --- Discussion --- p.67References --- p.7