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Publicação
Dissecting how LAMA2 modulation impact melanoma progression in a 3D in vitro model
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Autores
Resumo(s)
Melanoma, although not the most prevalent form of skin cancer, is the most aggressive and fatal. Melanoma progression and aggressiveness are dependent not only on the inherent characteristics and behavior of cancer cells, but also on the tumor microenvironment (TME), which surrounds cancer cells and is mainly composed by extracellular matrix (ECM), fibroblasts and immune cells. ECM is a key driver of tumor progression and changes in its compositions and organization can directly influence cancer cell behavior. This project aimed to understand how modifications in ECM components, particularly in the laminin α2-chain (encoded by the LAMA2 gene) affects melanoma cells, regarding DNA damage, growth, and migration. Melanoma cells (A375 cell line) with LAMA2 deletion (LAMA2 KO) were used and DNA damage, growth and their migration capacity, were compared with wild-type (WT) melanoma cells, using both 2D and 3D models (spheroids). Previous data from the host laboratory indicated that A375 LAMA2 KO cells displayed higher levels of phosphorylated p53. Contrasting, here a decreased in the phosphorylated p53 levels in LAMA2 KO spheroids with 10 days in culture was observed. Differences were also detected regarding the area of spheroids, with a significant increase in the area of LAMA2 KO spheroids 20 days after being established. Migration and RAC1 expression, a regulator of cell migration, were decreased in both 2D and 3D models in LAMA2 KO cells in comparison to wildtype counterparts. In addition, this project aimed to establish a co-culture system of melanoma and fibroblasts, which was successfully achieved. Collectively, this work advances our understanding of the critical role of the laminin-α2 chain in melanoma and establishes essential assays for characterizing spheroid cultures which more closely mimic the in vivo environment. These improved models provide a valuable platform to study tumor biology and microenvironmental interactions with greater physiological relevance.
Descrição
Tese de mestrado, Biologia do Organismo e Evolução , 2025, Universidade de Lisboa, Faculdade de Ciências
Palavras-chave
Melanoma Extracellular matrix LAMA2 Spheroids Co-culture