Thesis
Targeting inhibitory kappa B kinase alpha (IKKα) signalling in glioblastoma
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2024
- Thesis identifier
- T16933
- Person Identifier (Local)
- 201860008
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- Introduction: Cancer is a broad term, which covers a number of conditions characterised by uncontrolled cellular proliferation mainly due to genetic mutations. One of the most aggressive types of cancer is Glioblastoma multiforme (GBM). Presently, glioblastoma is considered an incurable type of cancer. Therefore, scientists are studying new patterns and mechanisms to supress GBM; one of which is through interrupting specific signalling pathways in tumour cells. One of the prominent pathways driving the tumour cell signalling responses and showing significant effects on cancer hallmarks, is the Nuclear Factor Kappa-B (NF-κB) pathway. Aim: The aim of the study is to investigate the pharmacological effect of selective IKKα inhibitors through examining their selectivity within both the non-canonical and canonical NF-κBpathways and their effect on IKKα-dependent cellular processes that underpin the phenotypic outcomes that support GBM development. Methods: Western Blotting was pursued with both whole cell extracts and crude nuclear extracts to determine the expression/phosphorylation/localisation of key signalling proteins. EMSA was used to determine the protein-DNA binding activity while MTT assay and clonogenic assay were used to identify cellular viability and clonogenic survival, respectively. All data shown were expressed as mean ± S.E.M. Statistical analysis was performed using GraphPad Prism version 10.1.0. The statistical significance of differences between mean values from control and treated groups were determined by one-way analysis of variance (ANOVA) with Dunnett’s post-test (p<0.05 was considered significant). Results: TWEAK and TNFα were used to stimulate T98G and/or UVW cells in increasing concentrations starting from 1 ng/ml up to 100 ng/ml or over time. Results showed that TWEAK was able to phosphorylate p100 in a concentrationdependent manner. 10 ng/ml of TWEAK were able stimulate p100 phosphorylation over time.It was also able to process p100 to p52/RelB and translocation to the nucleus in T98G GBM cells. TNFα was, to a lesser extent compared to TWEAK, able to phosphorylate p100 in a concentration-dependent manner. 10 ng/ml of TNFα were able to stimulate p100 phosphorylation over time. It was also able to process p100 to p52/RelB and translocation to the nucleus of T98G GBM cells. SU1433 and SU1644, were both able to inhibit, in a concentration-dependent manner, TWEAK-stimulated phosphorylation of p100 as well as p52 processing and nuclear translocation. Furthermore, they affected the phenotypic characteristics of GBM cell lines as well as protein-DNA binding activity. Conclusion: TWEAK can activate the non-canonical NF-κB pathway in glioblastoma cell lines in a concentration-dependent manner as well as over time. Moreover, SU1433 and SU1644 can inhibit TWEAK-stimulated non-canonical NF-κB pathway activation in Glioblastoma cell lines in a concentration dependent manner. These compounds can, hence, affect the phenotypic characteristics of GBM cells and could be potentially developed as drugs for the future management of brain cancer.
- Advisor / supervisor
- Paul, Andrew
- Boyd, Marie
- Resource Type
- DOI
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PDF of thesis T16933 | 2024-05-21 | Public | Download |