Fibroblasts potentiate melanoma cells in vitro invasiveness induced by UV-irradiated keratinocytes

Abstract

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/cm 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.

Single-Cell RNA Sequencing Unravels Heterogeneity of the Stromal Niche in Cutaneous Melanoma Heterogeneous Spheroids

“…Beside immune cells, CAFs produce numerous factors that stimulate tumour growth [42]. Among factors that enhance cancer growth and spreading, both IL-6 and IL-8 play a fundamental role not only in cutaneous but also in uveal melanoma [15,[43][44][45][46]. This observation can demonstrate how the model conditions can mimic the microenvironment.…”

“…This phenotype is triggered in the dermis by either extrinsic stimuli (oxidative stress, UV-related DNA damage), or intrinsic factors of chronological ageing. Fibroblasts are consequently switched into pro-inflammatory cells, acquiring the ability to affect adjacent cells of the local microenvironment [15]. Senescent fibroblasts produce numerous factors, e.g., cytokines, chemokines, growth factors or extracellular matrix proteins [9].…”

Heme oxygenase 1 facilitates cell proliferation via the B-Raf-ERK signaling pathway in melanoma

“…Melanomas arise from the pigment-producing cells called melanocytes, which normally reside in the epiderma and extend their dendrites into keratinocytes for melanin transportation and other cell-cell contacts [4]. Melanocytes also interact with skin fibroblasts and basal cells [17]. Thus, many researchers believe that the microenvironment of melanocytes plays an important role in melanoma development [18].…”

A new clinical tool to predict outcome in early-stage melanoma patients

“…IL-4 is the most important Th2 cytokine, and it is mainly produced by activated T cells, mast cells, basophils and eosinophils in order to regulate lymphocyte proliferation and survival 37 . Interestingly, elevated serum IL-6 was correlated with a poor prognosis in melanoma, while IL-4 is thought to promote the proliferation and survival of several cancer cells 39,40,41 . In line with previous findings, here early stage (I or II) melanoma patients that developed metastasis had significantly higher levels of serum IL-4…”