Uveal melanoma cells elicit retinal pericyte phenotypical and biochemical changes in an in vitro model of coculture
Autor: | Alfio Distefano, Mario Salmeri, Gabriella Lupo, Anna Longo, Angela Maria Amorini, Guido Zanghì, Giovanni Giurdanella, Carmelina Daniela Anfuso, Cesarina Giallongo |
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Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
Uveal Neoplasms
Becaplermin Vimentin lcsh:Chemistry STAT3 Uveal melanoma Cancer-Associated Fibroblasts Cell Movement Tumor Microenvironment Melanoma lcsh:QH301-705.5 Spectroscopy biology Chemistry General Medicine PDGF-B Neoplasm Proteins Computer Science Applications Gene Expression Regulation Neoplastic Interleukin 10 medicine.anatomical_structure Matrix Metalloproteinase 9 Imatinib Mesylate Pericyte Platelet-derived growth factor receptor Motility Article Retina Catalysis Receptor Platelet-Derived Growth Factor beta Transforming Growth Factor beta1 Inorganic Chemistry Gentamicin protection assay medicine Humans Physical and Theoretical Chemistry Molecular Biology Cell Proliferation Wound Healing Tumor microenvironment Organic Chemistry medicine.disease Coculture Techniques Coculture lcsh:Biology (General) lcsh:QD1-999 biology.protein Cancer research Pericytes |
Zdroj: | International Journal of Molecular Sciences Volume 21 Issue 15 International Journal of Molecular Sciences, Vol 21, Iss 5557, p 5557 (2020) |
Popis: | Vascular pericytes are an important cellular component in the tumor microenvironment, however, their role in supporting cancer invasion is poorly understood. We hypothesized that PDGF-BB could be involved in the transition of human retinal pericytes (HRPC) in cancer-activated fibroblasts (CAF), induced by the 92.1 uveal melanoma (UM) cell line. In our model system, HRPC were conditioned by co-culturing with 92.1UM for 6 days (cHRPC), in the presence or absence of imatinib, to block PDGF receptor-&beta (PDGFR&beta ). The effects of the treatments were tested by wound healing assay, proliferation assay, RT-PCR, high-content screening, Western blot analysis, and invasion assay. Results showed profound changes in cHRPC shape, with increased proliferation and motility, reduction of NG2 and increase of TGF-&beta 1, &alpha SMA, vimentin, and FSP-1 protein levels, modulation of PDGF isoform mRNA levels, phospho-PDGFR&beta and PDGFR&beta as well as phospho-STAT3 increases. A reduction of IL-1&beta and IFN&gamma and an increase in TNF&alpha IL10, and TGF-&beta 1, CXCL11, CCL18, and VEGF mRNA in cHRPC were found. Imatinib was effective in preventing all the 92.1UM-induced changes. Moreover, cHRPC elicited a significant increase of 92.1UM cell invasion and active MMP9 protein levels. Our data suggest that retinal microvascular pericytes could promote 92.1UM growth through the acquisition of the CAF phenotype. |
Databáze: | OpenAIRE |
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