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The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis
Bezuidenhout, L. W. 2025. The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis. Unpublished masters thesis. Ste...
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Stellenbosch : Stellenbosch University
2025
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| _version_ | 1867613875638108160 |
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| access_status_str | Open Access |
| author | Bezuidenhout, Ludick Wynand |
| author2 | Baker, Bienyameen |
| author_browse | Baker, Bienyameen Bezuidenhout, Ludick Wynand |
| author_facet | Baker, Bienyameen Bezuidenhout, Ludick Wynand |
| author_sort | Bezuidenhout, Ludick Wynand |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Bezuidenhout, L. W. 2025. The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain
containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis. Unpublished masters thesis. Stellenbosch: Stellenbosch University [online]. Available: https://scholar.sun.ac.za/items/4f171045-c058-445b-a861-4ee27eb04c19 |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/132069 |
| institution | Stellenbosch University (South Africa) |
| last_indexed | 2026-06-10T12:43:06.129Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/132069 The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis Bezuidenhout, Ludick Wynand Baker, Bienyameen Stellenbosch University. Faculty of Medicine and Health Sciences. Dept. of Biomedical Sciences. Molecular Biology and Human genetics. Breast — Cancer — Research Cancer cells — Migration Molecular oncology UCTD Bezuidenhout, L. W. 2025. The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis. Unpublished masters thesis. Stellenbosch: Stellenbosch University [online]. Available: https://scholar.sun.ac.za/items/4f171045-c058-445b-a861-4ee27eb04c19 Thesis (MSc)--Stellenbosch University, 2025. ENGLISH ABSTRACT: Breast cancer is the most prevalent kind of cancer among females in the world. The term represents a highly heterogenous disease that with substantial variation between patients. The rate of decline in mortality has slowed in recent years for various reasons, but it may be addressed by the development of novel therapies that may be used in conjunction with traditional chemotherapy and radiation treatment. Host directed therapies can occupy this niche by modulating the host immune response to cancer. A key contributor to the morbidity of cancer is the metastatic phenotype, whereby cancer cells spread from their point of origin to other distant tissues in the body where they proliferate in secondary colonies. The list of proteins associated with cancer progression is ever growing, but recent studies have identified that the interferon induced protein with tetratricopeptide repeats 2(IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) proteins may act as inhibitors of the migratory capacity of cancer cells. In order to study the effect of IFIT2 and HERC5 overexpression on breast cancer cell lines, they were transfected with pcDNA3.1(+) mammalian expression vector for overexpression of the respective genes. Additionally, cells were treated with IFIT2 mRNA carrying lipid nanoparticles (LNP). Cell migration was measured by mitomycin C (MMC) assays. WST-1 cell proliferation assays were performed in order to test for any apoptotic effect that may be present. The results of the study showed that MDA-MB-231 triple negative breast cancer cells were not impeded in terms of cell migration by plasmid mediated IFIT2 overexpression. Nor was there any evidence that IFIT2 induced apoptosis to any significant degree. MCF7 cells were significantly affected by IFIT2 overexpression. The Mission siRNA transfection reagent used in the transfection protocols was also found to have a significant effect on cell migration. To such an extent that it cannot be said for certain which component of the treatment, the IFIT2 overexpression or the transfection reagent, is more responsible for limiting cancer cell migration. LNP treatments did not result in significant impediment to cell migration in either cell line at tested concentrations, in contrast to IFIT2 plasmid treatments which remained effective in MCF7 cells in a direct comparison. In conclusion, due to limited replicates it cannot be said with certainty that IFIT2 is ineffective in limiting MDA-MB-231 cell migration. MCF7 cells do display sensitivity to treatments, though the unexpected activity of the siRNA transfection reagent obfuscates the component responsible for the phenomena. We additionally found no evidence that IFIT2 or HERC5 induced apoptosis in either cell line. AFRIKAANSE OPSOMMING: Borskanker is die mees algemene soort kanker onder vroue wêreldwyd. Die term verwys na 'n hoogs heterogene siekte met aansienlike variasie tussen pasiënte. Die tempo van sterftesyferafname het in onlangse jare verlangsaam weens verskeie redes, maar dit kan aangespreek word deur die ontwikkeling van nuwe terapieë wat saam met tradisionele chemoterapie en bestralingsterapie gebruik kan word. Gasheergerigte terapieë kan hierdie nis vul deur die gasheer se immuunrespons teen kanker te moduleer. ’n Belangrike bydraer tot die morbiditeit van kanker is die metastatiese fenotipe, waar kankerselle vanaf hul oorspronklike punt versprei na ander afgeleë weefsels in die liggaam, waar hulle in sekondêre kolonies vermeerder. Die lys proteïene wat met kankerontewikkeling geassosieer word, groei voortdurend, maar onlangse studies het geïdentifiseer dat die interferon-geïnduseerde proteïen met tetratrikopeptiedherhalings 2 (IFIT2) en HECT- en RLD-domeinbevattende E3-ubiquitienproteïenligase 5 (HERC5) proteïene moontlik as inhibeerders van die migrasie-vermoë van kankerselle kan optree Om die effek van IFIT2 en HERC5 op borskankersellyne te bestudeer, is die selle getransfekteer met die pcDNA3.1(+) eukariotiese uitdrukkingsvektor om ooruitdrukking van die onderskeie gene te bewerkstellig. Verder is die selle behandel met IFIT2 mRNA wat deur lipied-nanopartikels (LNP) gedra word. Kankersel migrasie is gemeet deur mitomisien C (MMC) toetse. WST-1 selproliferasie toetse is uitgevoer om enige apoptotiese effek wat moontlik teenwoordig is, te evalueer. Die resultate van die studie het getoon dat MDA-MB-231 borskankerselle nie belemmer is in terme van selmigrasie deur plasmied-gemedieerde IFIT2-ooruitdrukking nie. Daar was ook geen bewys dat IFIT2 apoptose tot 'n beduidende mate geïnduseer het nie. MCF7-selle is egter duidelik beïnvloed deur IFIT2-ooruitdrukking. Daar is ook bevind dat die Mission siRNA-transfeksie-reagens, wat in die transfeksieprotokolle gebruik is, 'n beduidende effek op selmigrasie gehad het. Tot so 'n mate dat dit nie met sekerheid gesê kan word watter komponent van die behandeling, naamlik IFIT2- ooruitdrukking of die transfeksie-reagens, meer verantwoordelik is vir die beperking van kankerselmigrasie nie. LNP-behandelings het nie tot 'n betekenisvolle belemmering van selmigrasie in enige van die twee selllyne by getoetste konsentrasies gelei nie, in teenstelling met IFIT2- plasmiedbehandelings, wat effektief gebly het in MCF7-selle. Ten slotte, as gevolg van 'n beperkte aantal herhalings, kan daar nie met sekerheid gesê word dat IFIT2 nie effektief is in die beperking van MDA-MB-231-selmigrasie nie. MCF7-selle toon wel sensitiwiteit teenoor die behandelings, hoewel die onverwagte aktiwiteit van die siRNA-transfeksie-reagens die identifikasie van die komponent wat vir die verskynsel verantwoordelik is, bemoeilik. Ons het ook geen bewys gevind dat IFIT2 of HERC5 apoptose in enige van die selllyne geïnduseer het nie. Masters 2025-05-22T07:54:09Z 2025-05-22T07:54:09Z 2025-03 Thesis https://scholar.sun.ac.za/handle/10019.1/132069 Stellenbosch University 54 pages : illustrations application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Breast — Cancer — Research Cancer cells — Migration Molecular oncology UCTD Bezuidenhout, Ludick Wynand The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis |
| title | The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis |
| title_full | The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis |
| title_fullStr | The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis |
| title_full_unstemmed | The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis |
| title_short | The anti-cancer effect of interferon induced protein with tetratricopeptide repeats 2 (IFIT2) and HECT and RLD domain containing E3 ubiquitin protein ligase 5 (HERC5) on breast cancer cell lines in the context of cell migration and apoptosis |
| title_sort | anti cancer effect of interferon induced protein with tetratricopeptide repeats 2 ifit2 and hect and rld domain containing e3 ubiquitin protein ligase 5 herc5 on breast cancer cell lines in the context of cell migration and apoptosis |
| topic | Breast — Cancer — Research Cancer cells — Migration Molecular oncology UCTD |
| url | https://scholar.sun.ac.za/handle/10019.1/132069 |
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