Browsing by Author "Stone, Marian"

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    Defining the functional role of MAGE-C1 in Multiple Myeloma
    (2024) Stone, Marian; Shires, Karen
    The expression of Cancer/Testis antigens (CTAs) in various cancers, including multiple myeloma, makes them attractive targets for therapy and biomarker development. Despite intensive studies, their potential has not been fully realized due to lack of knowledge regarding their pathogenic role. To start elucidating the function of melanoma antigen (MAGE) protein-C1 (MAGE-C1) in multiple myeloma, this project aimed to firstly establish the suitability of culture cell line RPMI-8226 as an in vitro multiple myeloma model. Additionally, the project aimed to establish relevant reagents and techniques to study MAGE proteins by comparing MAGE-C1 to better described CTAs: MAGE-C2 and MAGE-A1. Firstly, the expression of MAGE-C1, MAGE-A1, MAGE-C2 and their respective binding partners, Ski interacting protein (SKIP) and Ring-box 1 (RBX1), was confirmed in RPMI-8662. Next, functional similarity was investigated through co-immunoprecipitation, to determine MAGE-C1 interaction with RBX1 or SKIP, and immunofluorescence microscopy, to assess co-localisation of these proteins. Lastly, cellular pathways influenced by MAGE-C2 were evaluated using heat shock to induce a stress response and activate the pathways of interest. The expression of MAGE-C1, MAGE-A1, MAGE-C2 and their binding partners were confirmed at the mRNA level. Unfortunately, interaction with their binding partners could not be demonstrated using co-immunoprecipitation, although immunofluorescence microscopy showed co-localisation of MAGE-C1 with both SKIP and RBX1. An abnormal stress response, via the E3 ligases known to interact with MAGE-C2 was observed in RPMI-8226. An array of reagents and techniques for future MAGE investigations were established during this study. However, the findings of this project indicated that RPMI-8226 may not be an ideal model for further MAGE investigations as initially hypothesized