Defining the functional role of MAGE-C1 in Multiple Myeloma
| dc.contributor.advisor | Shires, Karen | |
| dc.contributor.author | Stone, Marian | |
| dc.date.accessioned | 2025-04-11T12:37:47Z | |
| dc.date.available | 2025-04-11T12:37:47Z | |
| dc.date.issued | 2024 | |
| dc.date.updated | 2025-04-07T07:46:50Z | |
| dc.description.abstract | 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 | |
| dc.identifier.apacitation | Stone, M. (2024). <i>Defining the functional role of MAGE-C1 in Multiple Myeloma</i>. (). University of Cape Town ,Faculty of Health Sciences ,Department of Clinical Laboratory Sciences. Retrieved from http://hdl.handle.net/11427/41387 | en_ZA |
| dc.identifier.chicagocitation | Stone, Marian. <i>"Defining the functional role of MAGE-C1 in Multiple Myeloma."</i> ., University of Cape Town ,Faculty of Health Sciences ,Department of Clinical Laboratory Sciences, 2024. http://hdl.handle.net/11427/41387 | en_ZA |
| dc.identifier.citation | Stone, M. 2024. Defining the functional role of MAGE-C1 in Multiple Myeloma. . University of Cape Town ,Faculty of Health Sciences ,Department of Clinical Laboratory Sciences. http://hdl.handle.net/11427/41387 | en_ZA |
| dc.identifier.ris | TY - Thesis / Dissertation AU - Stone, Marian AB - 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 DA - 2024 DB - OpenUCT DP - University of Cape Town KW - Medicine LK - https://open.uct.ac.za PB - University of Cape Town PY - 2024 T1 - Defining the functional role of MAGE-C1 in Multiple Myeloma TI - Defining the functional role of MAGE-C1 in Multiple Myeloma UR - http://hdl.handle.net/11427/41387 ER - | en_ZA |
| dc.identifier.uri | http://hdl.handle.net/11427/41387 | |
| dc.identifier.vancouvercitation | Stone M. Defining the functional role of MAGE-C1 in Multiple Myeloma. []. University of Cape Town ,Faculty of Health Sciences ,Department of Clinical Laboratory Sciences, 2024 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/41387 | en_ZA |
| dc.language.iso | en | |
| dc.language.rfc3066 | eng | |
| dc.publisher.department | Department of Clinical Laboratory Sciences | |
| dc.publisher.faculty | Faculty of Health Sciences | |
| dc.publisher.institution | University of Cape Town | |
| dc.subject | Medicine | |
| dc.title | Defining the functional role of MAGE-C1 in Multiple Myeloma | |
| dc.type | Thesis / Dissertation | |
| dc.type.qualificationlevel | Masters | |
| dc.type.qualificationlevel | MSc |