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Title: Development of novel technologies used to measure tumour necrosis factor receptor signalling in living cells
Author: Tucker, S. J.
Awarding Body: University of Aberdeen
Current Institution: University of Aberdeen
Date of Award: 2004
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The aim of this work was firstly to assess novel reporter constructs as potential methods for investigating TNF signalling. The key early findings of this study suggested the GFP-linked reporter construct system as more suitable for studying TNF signalling. Subsequently a series of optimisation experiments maximised this system in HeLa and TF-1 cells. In both cell lines TNF strongly induced p38 MAPK, p42/44 MAPK, JNK and NF-κB reporter constructs. In functional terms p42/44 MAPK activation was shown to be critical for TF-1 cell proliferation, whilst JNK related to TNF-induced cell death in both cell types. Interestingly, p38 MAPK was also related to TNF-induced cell death, but only in TF-1 cells. In contrast, NF-κB were shown to exist in a co-inhibitory relationship, in which inhibition of one resulted in potentiation of the other. This was shown to be an important regulatory controller of cell death following TNF stimulation of HeLa and TF-1 cells. In addition, TNF-induced JNK activation was related to caspases, with suppression of caspases reducing the degree of JNK activity. Finally, receptor constructs were used to assess the signalling prowess of sodium salicylate in TF-1 cells, which activated the same signal pattern as TNF in dying TF-1 cells. Indeed sodium salicylate reciprocated the selective induction of cell death in proliferating TF-1 cells seen with TNF, suggesting potential for this drug in the treatment of leukaemia. Associated with early experiments, polymeric sponge toxin fractions (polyAPS and halitoxin) were shown to mediate successful HEK 293 and HeLa transfection on account of their porative activity.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available