TNF-alpha and metalloproteases as key players in melanoma cells aggressiveness

Stefania Rossi,Cinzia Senatore,Alessandro Weisz,Daniela D’Arcangelo,Antonio Facchiano,Claudio Tabolacci,Giovanni Nassa,Roberta Magliozzi,Paolo Pagnotto,Annamaria Salvati,Martina Cordella,Francesco Facchiano

doi:10.1186/s13046-018-0982-1

Abstract

BackgroundMelanoma aggressiveness determines its growth and metastatic potential. This study aimed at identifying new molecular pathways controlling melanoma cell malignancy.MethodsTen metastatic melanoma cell lines were characterized by their proliferation, migration and invasion capabilities. The most representative cells were also characterized by spheroid formation assay, gene- and protein- expression profiling as well as cytokines secretion and the most relevant pathways identified through bioinformatic analysis were tested by in silico transcriptomic validation on datasets generated from biopsies specimens of melanoma patients. Further, matrix metalloproteases (MMPs) activity was tested by zymography assays and TNF-alpha role was validated by anti-TNF cell-treatment.ResultsAn aggressiveness score (here named Melanoma AGgressiveness Score: MAGS) was calculated by measuring proliferation, migration, invasion and cell-doubling time in10human melanoma cell lines which were clustered in two distinct groups, according to the corresponding MAGS. SK-MEL-28 and A375 cell lines were selected as representative models for the less and the most aggressive phenotype, respectively. Gene-expression and protein expression data were collected for SK-MEL-28 and A375 cells by Illumina-, multiplex x-MAP-and mass-spectrometry technology. The collected data were subjected to an integrated Ingenuity Pathway Analysis, which highlighted that cytokine/chemokine secretion, as well as Cell-To-Cell Signaling and Interaction functions as well as matrix metalloproteases activity were significantly different in these two cell types. The key role of these pathways was then confirmed by functional validation. TNF role was confirmed by exposing cells to the anti-TNF Infliximab antibody. Upon such treatment melanoma cells aggressiveness was strongly reduced. Metalloproteases activity was assayed, and their role was confirmed by comparing transcriptomic data from cutaneous melanoma patients (n = 45) and benign nevi (n = 18).ConclusionsInflammatory signals such as TNF and MMP-2 activity are key intrinsic players to determine melanoma cells aggressiveness suggesting new venue sin the identification of novel molecular targets with potential therapeutic relevance.

Highlights

Melanoma aggressiveness determines its growth and metastatic potential
matrix metalloproteases (MMPs)-9 activation was associated with cancer growth and dissemination [12], its role in cutaneous melanoma was reported and its activation, mediated by NF-κB, was associated with the BRAFV600E mutation status [13]
Cutaneous melanoma (CM) aggressiveness has been associated with its mutational state, and with the anatomical site where the primary tumor occurs or, in addition, with the immunological status of patients observed in different cases of patients receiving immunosuppressive therapies [27, 28]

Summary

Introduction

Melanoma aggressiveness determines its growth and metastatic potential. This study aimed at identifying new molecular pathways controlling melanoma cell malignancy. MMP-9 activation was associated with cancer growth and dissemination [12], its role in cutaneous melanoma was reported and its activation, mediated by NF-κB, was associated with the BRAFV600E mutation status [13]. Another recent study reports the correlation of MMP-9 hypermethylation with its overexpression in melanoma [14] indicating novel molecular mechanisms underlying the MMPs activity and their modulatory role in melanoma aggressiveness

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