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Defining the functional role of MAGE-C1 in Multiple Myeloma
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 sta...
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| Format: | Thesis |
| Language: | English English |
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Department of Clinical Laboratory Sciences
2025
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| _version_ | 1867613336943722496 |
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| access_status_str | Open Access |
| author | Stone, Marian |
| author2 | Shires, Karen |
| author_browse | Shires, Karen Stone, Marian |
| author_facet | Shires, Karen Stone, Marian |
| author_sort | Stone, Marian |
| collection | Thesis |
| description | 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 |
| format | Thesis |
| id | oai:open.uct.ac.za:11427/41387 |
| institution | University of Cape Town (South Africa) |
| language | English eng |
| last_indexed | 2026-06-10T12:34:32.198Z |
| license_str | Not specified — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Department of Clinical Laboratory Sciences |
| publisherStr | Department of Clinical Laboratory Sciences |
| record_format | dspace |
| source_str | UCTD — University of Cape Town Open Access Repository |
| spelling | oai:open.uct.ac.za:11427/41387 Defining the functional role of MAGE-C1 in Multiple Myeloma Stone, Marian Shires, Karen Medicine 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 2025-04-11T12:37:47Z 2025-04-11T12:37:47Z 2024 2025-04-07T07:46:50Z Thesis / Dissertation Masters MSc http://hdl.handle.net/11427/41387 en eng application/pdf Department of Clinical Laboratory Sciences Faculty of Health Sciences University of Cape Town |
| spellingShingle | Medicine Stone, Marian Defining the functional role of MAGE-C1 in Multiple Myeloma |
| thesis_degree_str | Master's |
| title | Defining the functional role of MAGE-C1 in Multiple Myeloma |
| title_full | Defining the functional role of MAGE-C1 in Multiple Myeloma |
| title_fullStr | Defining the functional role of MAGE-C1 in Multiple Myeloma |
| title_full_unstemmed | Defining the functional role of MAGE-C1 in Multiple Myeloma |
| title_short | Defining the functional role of MAGE-C1 in Multiple Myeloma |
| title_sort | defining the functional role of mage c1 in multiple myeloma |
| topic | Medicine |
| url | http://hdl.handle.net/11427/41387 |
| work_keys_str_mv | AT stonemarian definingthefunctionalroleofmagec1inmultiplemyeloma |