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.

Biotechnology and Biomedicine Center of the Academy of Sciences and Charles University in Vestec Průmyslová 595 Vestec u Prahy Prague Czech Republic

Cell and Experimental Pathology Department of Translational Medicine Lund University Clinical Research Centre Skåne University Hospital Jan Waldenströms gata 35 21421 Malmö Sweden

Department of Cell Biology Faculty of Sciences Charles University Prague Viničná 7 120 00 Prague 2 Czech Republic

Department of Dermatovenereology 1st Faculty of Medicine Charles University U Nemocnice 2 Prague 2 Czech Republic

Department of Otorhinolaryngology Head and Neck Surgery 1st Faculty of Medicine Charles University 5 Úvalu 5 Prague 5 Czech Republic

Institute of Anatomy 1st Faculty of Medicine Charles University U Nemocnice 3 Prague 2 Czech Republic

Institute of Biochemistry and Experimental Oncology 1st Faculty of Medicine Charles University U Nemocnice 5 Prague 2 Czech Republic

Institute of Molecular Genetics Academy of Sciences of the Czech Republic vvi Vídeňská 1083 Prague 4 Czech Republic

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