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

S. Rossi, M. Cordella, C. Tabolacci, G. Nassa, D. D'Arcangelo, C. Senatore, P. Pagnotto, R. Magliozzi, A. Salvati, A. Weisz, A. Facchiano, F. Facchiano

Research output: Contribution to journalArticlepeer-review

Abstract

Background: Melanoma aggressiveness determines its growth and metastatic potential. This study aimed at identifying new molecular pathways controlling melanoma cell malignancy. Methods: Ten 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. Results: An 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). Conclusions: Inflammatory 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. © 2018 The Author(s).
Original languageEnglish
JournalJournal of Experimental and Clinical Cancer Research
Volume37
Issue number1
DOIs
Publication statusPublished - 2018

Keywords

  • Cancer
  • Cutaneous melanoma
  • Cytokines
  • Inflammation
  • Malignancy
  • Metalloproteases
  • Proteomics
  • TNF
  • Uveal melanoma
  • eotaxin
  • gamma interferon
  • gelatinase A
  • granulocyte colony stimulating factor
  • infliximab
  • interleukin 1 receptor blocking agent
  • interleukin 10
  • interleukin 12
  • interleukin 1beta
  • interleukin 6
  • interleukin 8
  • interleukin 9
  • matrix metalloproteinase
  • platelet derived growth factor BB
  • RANTES
  • tumor necrosis factor
  • vasculotropin
  • cytokine
  • metalloproteinase
  • A-375 cell line
  • Article
  • assay
  • bioinformatics
  • cell interaction
  • cell invasion
  • cell migration
  • cell proliferation
  • computer model
  • controlled study
  • cytokine release
  • enzyme activity
  • gene expression
  • human
  • human cell
  • intracellular signaling
  • major clinical study
  • mass spectrometry
  • melanoma
  • Melanoma Aggressiveness Score
  • priority journal
  • protein expression
  • scoring system
  • SK-MEL-28 cell line
  • spheroid formation assay
  • transcriptomics
  • zymography
  • genetics
  • metabolism
  • pathology
  • physiology
  • proteomics
  • skin tumor
  • tumor cell line
  • Cell Line, Tumor
  • Cell Proliferation
  • Humans
  • Melanoma
  • Skin Neoplasms
  • Tumor Necrosis Factor-alpha

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