Fibroblasts potentiate melanoma cells in vitro invasiveness induced by UV-irradiated keratinocytes
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Melanoma represents a malignant disease with steadily increasing incidence. UV-irradiation is a recognized key factor in melanoma initiation. Therefore, the efficient prevention of UV tissue damage bears a critical potential for melanoma prevention. In this study, we tested the effect of UV irradiation of normal keratinocytes and their consequent interaction with normal and cancer-associated fibroblasts isolated from melanoma, respectively. Using this model of UV influenced microenvironment, we measured melanoma cell migration in 3-D collagen gels. These interactions were studied using DNA microarray technology, immunofluorescence staining, single cell electrophoresis assay, viability (dead/life) cell detection methods, and migration analysis. We observed that three 10 mJ/cm2 fractions at equal intervals over 72 h applied on keratinocytes lead to a 50% increase (p < 0.05) in in vitro invasion of melanoma cells. The introduction cancer-associated fibroblasts to such model further significantly stimulated melanoma cells in vitro invasiveness to a higher extent than normal fibroblasts. A panel of candidate gene products responsible for facilitation of melanoma cells invasion was defined with emphasis on IL-6, IL-8, and CXCL-1. In conclusion, this study demonstrates a synergistic effect between cancer microenvironment and UV irradiation in melanoma invasiveness under in vitro condition.
KeywordsCancer-associated fibroblasts Keratinocytes Cancer microenvironment Cytokine Chemokine Melanoma
This publication is a result of the project implementation: “The equipment for metabolomic and cell analyses”, registration number CZ.1.05/2.1.00/19.0400, supported by Research and Development for Innovations Operational Programme (RDIOP) co-financed by European regional development fund and the state budget of the Czech Republic. This study was also supported by the Grant Agency of the Czech Republic (Project no. 16-05534S), AZV 16-29032A, the Charles University (project of Specific University Research, GAUK 165015 and PROGRESS 28 and UNCE 23014) and by the Ministry of Education, Youth and Sports of CR within the National Sustainability Program II (Project BIOCEV-FAR reg. no. LQ1604), and by the project BIOCEV (CZ.1.05/1.1.00/02.0109). The part of the study was performed by the equipment for metabolomics and cell analyses (Grant no. CZ.1.05/2.1.00/19.0400) supported by the Research and Development for Innovations Operational Program, co-financed by the European regional development fund and the state budget of the Czech Republic.
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