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Apoptosis of tumor cells : role of TRAIL and caspase 10
Truxová, Iva ; Živný, Jan (advisor) ; Anděra, Ladislav (referee)
One of the key features of cancer cells is the ability to escape programmed cell death (apoptosis). As a mechanism of apoptosis inactivation in cancer cells, somatic mutations of pro-apoptotic genes have been reported in many cancers. Caspase 10 is an initiator caspase whose physiological function remains poorly understood. Also the ability of caspase 10 to substitute for caspase 8 in the death receptors apoptotic pathway is still controversial. However, the fact that some of the mutations found in CASP10 gene was associated with apoptosis defects (79, 81) suggest that caspase 10 could be also important in apoptosis initiation. In our lab, there was found a heterozygous mutation in CASP10 gene of Jurkat (human T-acute lymphoblastic leukemia) clone resistant to TRAIL (J-TR1). This mutation influence the amino acid composition close to the active site of the enzyme. The aim of this thesis was to confirm the mutation by ARMS-PCR and to determine if an overexpression of normal (unmutated) or mutated caspase 10 D in TRAIL sensitive and/or TRAIL resistant Jurkat cells (J-WT and/or J-TR1) will influence TRAIL induced apoptosis. Mutation was confirmed. We created J-WT and J-TR1 stable clones transfected by vector with unmutated or mutated CASP10 D (CASP10 D WT or CASP10 D MUT). CASP10 D MUT overexpression in J-WT...
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Immunogenic cell death and it's relevance for biology and therapy of malignant diseases
Truxová, Iva ; Špíšek, Radek (advisor) ; Říhová, Blanka (referee) ; Kalina, Tomáš (referee)
Immunostimulatory potential of tumor cells depends on various factors, including primarily tumor antigen repertoire and the capacity to emit molecules associated with cellular stress or injury, so called DAMPs, during immunogenic forms of cell death. These molecules mainly act on dendritic cells (DCs), thus activating the antitumor immune response. Several immunogenic cell death (ICD) inducers have been described in the past years. The contribution of my PhD thesis into this topic was the characterization of the apoptotic pathways activated by high hydrostatic pressure (HHP). HHP induces rapid tumor cell death accompanied by DAMP release (mainly calreticulin (CRT), HSP70, HSP90, HMGB1 and ATP) that is characterized by the overproduction of reactive oxygen species (ROS) causing the establishment of integrated stress response. ROS-PERK-eIF2α-caspase-2-caspase-8 signaling pathway plays an essential role in CRT translocation to the tumor cell surface upon HHP treatment, thus influencing the immunogenic potential of these cells. Moreover, the importance of ICD concept was also confirmed in vivo. The results point out that the presence of CRT on the surface of malignant blasts from AML patients correlates with the activation of specific antitumor immune response and improved clinical outcome. Another...
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Apoptosis of tumor cells : role of TRAIL and caspase 10
Truxová, Iva ; Živný, Jan (advisor) ; Anděra, Ladislav (referee)
One of the key features of cancer cells is the ability to escape programmed cell death (apoptosis). As a mechanism of apoptosis inactivation in cancer cells, somatic mutations of pro-apoptotic genes have been reported in many cancers. Caspase 10 is an initiator caspase whose physiological function remains poorly understood. Also the ability of caspase 10 to substitute for caspase 8 in the death receptors apoptotic pathway is still controversial. However, the fact that some of the mutations found in CASP10 gene was associated with apoptosis defects (79, 81) suggest that caspase 10 could be also important in apoptosis initiation. In our lab, there was found a heterozygous mutation in CASP10 gene of Jurkat (human T-acute lymphoblastic leukemia) clone resistant to TRAIL (J-TR1). This mutation influence the amino acid composition close to the active site of the enzyme. The aim of this thesis was to confirm the mutation by ARMS-PCR and to determine if an overexpression of normal (unmutated) or mutated caspase 10 D in TRAIL sensitive and/or TRAIL resistant Jurkat cells (J-WT and/or J-TR1) will influence TRAIL induced apoptosis. Mutation was confirmed. We created J-WT and J-TR1 stable clones transfected by vector with unmutated or mutated CASP10 D (CASP10 D WT or CASP10 D MUT). CASP10 D MUT overexpression in J-WT...
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