Sphingosine-1-phosphate induces cytokine expression, migration and MMPs secretion in melanoma cells: role of Filamin A

CAMPOS LUDMILA ESTEFANÍA; MORELLATTO RUGGIERI LUCIANA; CASTRO MELINA G; RODRIGUEZ YAMILA I; ALVAREZ SERGIO E

Mar del Plata

Congreso; LXI REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE INVESTIGACIÓN CLÍNICA (SAIC), LXIV REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE INMUNOLOGÍA (SAI) XLVIII REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE FARMACOLOGÍA EXPERIMENTAL (SAFE); 2016

SAIC, SAI y SAFE

Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid linked to chronic inflammation and cancer. Once produced intracellularly by two sphingosine kinases SK1 and SK2, S1P might act as a) a second messenger or b) as autocrine/paracrine metabolite by acting through G protein-coupled receptors (GPCRs), named S1PR1-5. Most of S1P actions are mediated by its receptors, including migration, invasion and angiogenesis. Previously, we have shown that extracellular S1P induces NF-κB activation in melanoma cell lines that do not express Filamin-A (FLNa), an actin binding protein. Here we explored some biological actions triggered by extracellular S1P in two melanoma cell lines: M2 cells (FLNa-) and A7 cells (FLNa+). First, we analyzed mRNA expression of NF-κB-regulated cytokines and chemokines by qPCR. Our results demonstrate that S1P stimulates mRNA expression of IL-6, TNFα, CCL2 and CCL5 only in M2 cells. By using Boyden?s chamber migration assays, we found that S1P induces migration in M2 cells but the effect is minor in A7 cells. Moreover, S1P protected from serum depletion-induced apoptosis in M2 cells. In concordance with these results, MTT experiments indicate that S1P preserve cell viability in M2 cells while its effect is less noticeable on A7 cells. In addition, gelatin degradation assays showed that S1P induces MMP9 secretion in M2 cells. Interestingly, a differential behavior is observed in A7 cells: S1P induced MMP2, but not MMP9, secretion, suggesting that FLNa may play a role in the regulation of MMPs release. In summary, our experiments indicate that FLNa expression modulates extracellular S1P induced-cytokine expression, MMP secretion, migration and proliferation in melanoma cells.