Publicação
Development of an in vitro model of mucinous tumors
| Resumo: | Mucinous carcinomas are tumors with poor prognosis and secrete a large amount of mucus. The development of an in vitro model that reconstitutes the mucin-rich microenvironment of mucinous carci- nomas can provide a better understanding on how the aberrant mucus production relates to the pro- gression of these tumors. Furthermore, it responds to the lack of predictive in vitro models and leads the way to the replacement of standard animal models. In this project, colorectal mucinous carcinomas are studied, for which several mucin-secreting cell lines are available. We demonstrate that crosslinked bovine submaxillary mucin gels function as a platform to encapsulate cancer cells and obtain spheroids that grow and proliferate over 10 days. More importantly, they resist 5-FU and the drug resistance was different between cell lines. Although at a slower rate than HT-29 MTX, LS174T cells reached equivalent spheroids diameter (156.5 and 187.9 μm, respectively) and metabolic activity (3- and 4- fold increases, respectively). LS174T also showed the greater physical barrier effect against 5-FU in monolayer (2D) or encapsulated (3D) in BSM, but also a potential biological barrier caused by cell entanglement in mucin-gel for 10 days. In the 2D model, LS174T cell viability went from 45.9 ± 8.81% without gel to 84.2 ± 1.96% with muc-gel covering the cells. When encapsulated and further challenged with 50.0 mM of 5-FU, 59.6 ± 7.56% cells were viable after 6 h, whereas 76.2 ± 5.20% of cells survived after 10 days of interaction with BSM. Thus, these results unlock an exciting path to apply mucins as a scaffold for mucinous cancer cells and enlighten necessary refinements to be implemented in future studies towards the ultimate goal of building a robust 3D model capable of recreating the in vivo tumor microenvironment and serving as a high-throughput platform for drug screening in the pharmaceutical industry. |
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| Autores principais: | Benavente, Ana Luísa da Silva Ribeiro |
| Assunto: | Colorectal cancer Drug-resistance Encapsulation Hydrogel Mucins Tumor Micro-environment |
| Ano: | 2022 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade Nova de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Institucional da UNL |
| Resumo: | Mucinous carcinomas are tumors with poor prognosis and secrete a large amount of mucus. The development of an in vitro model that reconstitutes the mucin-rich microenvironment of mucinous carci- nomas can provide a better understanding on how the aberrant mucus production relates to the pro- gression of these tumors. Furthermore, it responds to the lack of predictive in vitro models and leads the way to the replacement of standard animal models. In this project, colorectal mucinous carcinomas are studied, for which several mucin-secreting cell lines are available. We demonstrate that crosslinked bovine submaxillary mucin gels function as a platform to encapsulate cancer cells and obtain spheroids that grow and proliferate over 10 days. More importantly, they resist 5-FU and the drug resistance was different between cell lines. Although at a slower rate than HT-29 MTX, LS174T cells reached equivalent spheroids diameter (156.5 and 187.9 μm, respectively) and metabolic activity (3- and 4- fold increases, respectively). LS174T also showed the greater physical barrier effect against 5-FU in monolayer (2D) or encapsulated (3D) in BSM, but also a potential biological barrier caused by cell entanglement in mucin-gel for 10 days. In the 2D model, LS174T cell viability went from 45.9 ± 8.81% without gel to 84.2 ± 1.96% with muc-gel covering the cells. When encapsulated and further challenged with 50.0 mM of 5-FU, 59.6 ± 7.56% cells were viable after 6 h, whereas 76.2 ± 5.20% of cells survived after 10 days of interaction with BSM. Thus, these results unlock an exciting path to apply mucins as a scaffold for mucinous cancer cells and enlighten necessary refinements to be implemented in future studies towards the ultimate goal of building a robust 3D model capable of recreating the in vivo tumor microenvironment and serving as a high-throughput platform for drug screening in the pharmaceutical industry. |
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