Generation and Characterization of a Tumor Stromal Microenvironment and Analysis of Its Interplay with Breast Cancer Cells: An In Vitro Model to Study Breast Cancer-Associated Fibroblast Inactivation.

Autor: Romano V; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., Ruocco MR; Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131 Naples, Italy., Carotenuto P; TIGEM, Telethon Institute of Genetics and Medicine, 80078 Naples, Italy.; Medical Genetics, Department of Translational Medical Science, University of Naples Federico II, 80131 Naples, Italy., Barbato A; TIGEM, Telethon Institute of Genetics and Medicine, 80078 Naples, Italy., Venuta A; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., Acampora V; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., De Lella S; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., Vigliar E; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., Iaccarino A; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., Troncone G; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., Calì G; IEOS Istituto di Endocrinologia e Oncologia Sperimentale 'G. Salvatore', National Council of Research, 80131 Naples, Italy., Insabato L; Anatomic Pathology Unit, Department of Advanced Biomedical Sciences, School of Medicine, University of Naples Federico II, 80131 Naples, Italy., Russo D; Anatomic Pathology Unit, Department of Advanced Biomedical Sciences, School of Medicine, University of Naples Federico II, 80131 Naples, Italy., Franco B; TIGEM, Telethon Institute of Genetics and Medicine, 80078 Naples, Italy.; Medical Genetics, Department of Translational Medical Science, University of Naples Federico II, 80131 Naples, Italy.; Scuola Superiore Meridionale, School for Advanced Studies, 80138 Naples, Italy., Masone S; Department of Clinical Medicine and Surgery, University of Naples Federico II, 80131 Naples, Italy., Velotti N; Department of Advanced Biochemical Sciences, University of Naples Federico II, 80131 Naples, Italy., Accurso A; Department of General, Oncological, Bariatric and Endocrine-Metabolic Surgery, University of Naples Federico II, 80131 Naples, Italy., Pellegrino T; DAI Chirurgia Generale, Endocrinologia, Ortopedia e Riabilitazione, Azienda Ospedaliera Universitaria Federico II, 80131 Naples, Italy., Fiume G; Department of Experimental and Clinical Medicine, University 'Magna Graecia' of Catanzaro, 88100 Catanzaro, Italy., Belviso I; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., Montagnani S; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., Avagliano A; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy., Arcucci A; Department of Public Health, University of Naples Federico II, 80131 Naples, Italy.
Jazyk: angličtina
Zdroj: International journal of molecular sciences [Int J Mol Sci] 2022 Jun 20; Vol. 23 (12). Date of Electronic Publication: 2022 Jun 20.
DOI: 10.3390/ijms23126875
Abstrakt: Breast cancer-associated fibroblasts (BCAFs), the most abundant non-cancer stromal cells of the breast tumor microenvironment (TME), dramatically sustain breast cancer (BC) progression by interacting with BC cells. BCAFs, as well as myofibroblasts, display an up regulation of activation and inflammation markers represented by α-smooth muscle actin (α-SMA) and cyclooxygenase 2 (COX-2). BCAF aggregates have been identified in the peripheral blood of metastatic BC patients. We generated an in vitro stromal model consisting of human primary BCAFs grown as monolayers or 3D cell aggregates, namely spheroids and reverted BCAFs, obtained from BCAF spheroids reverted to 2D cell adhesion growth after 216 h of 3D culture. We firstly evaluated the state of activation and inflammation and the mesenchymal status of the BCAF monolayers, BCAF spheroids and reverted BCAFs. Then, we analyzed the MCF-7 cell viability and migration following treatment with conditioned media from the different BCAF cultures. After 216 h of 3D culture, the BCAFs acquired an inactivated phenotype, associated with a significant reduction in α-SMA and COX-2 protein expression. The deactivation of the BCAF spheroids at 216 h was further confirmed by the cytostatic effect exerted by their conditioned medium on MCF-7 cells. Interestingly, the reverted BCAFs also retained a less activated phenotype as indicated by α-SMA protein expression reduction. Furthermore, the reverted BCAFs exhibited a reduced pro-tumor phenotype as indicated by the anti-migratory effect exerted by their conditioned medium on MCF-7 cells. The deactivation of BCAFs without drug treatment is possible and leads to a reduced capability of BCAFs to sustain BC progression in vitro. Consequently, this study could be a starting point to develop new therapeutic strategies targeting BCAFs and their interactions with cancer cells.
Databáze: MEDLINE
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