Measurement of 45 cytokine, chemokine and growth factors in established cell culture supernatants and autologous serum from advanced melanoma patients

Abstract

Melanoma is one of the most aggressive forms of human cancer and its incidence has significantly increased worldwide over the last decades. This neoplasia has been characterized by the release of a wide variety of soluble factors, which could stimulate tumor cell proliferation and survival in an autocrine and paracrine manner. Consequently, we sought to evaluate the pattern of soluble factors produced by pre-metastatic and metastatic melanoma established cultures, and to determine whether these factors can be detected in the autologous serum of malignant melanoma patients. Our results showed that both melanoma cultures had a common profile of 27 soluble factors mainly characterized by the high expression of VEGF-A, IL-6, MCP-1, IL-8, and SDF-1. In addition, when we compared supernatants, we observed significant differences in VEGF-A, BDNF, FGF-2, and NGF-β concentrations. As we found in melanoma cultures, serum samples also had their specific production pattern composed by 21 soluble factors. Surprisingly, PDGF-BB and EGF were only found in serum, whereas IL-2, IL-4, IL-8, IL31, FGF2, and GRO-α were only expressed in the supernatant. Significant differences in PDGF-BB, MIP-1β, HGF, PIGF-1, BDNF, EGF, Eotaxin, and IP-10 were also found after comparing autologous serum with healthy controls. According to this, no correlation was found between culture supernatants and autologous serum samples, which suggests that some factors may act locally, and others systemically. Nonetheless, after validation of our results in an independent cohort of patients, we concluded that PDGF-BB, VEGF-A, and IP-10 serum levels could be used to monitor different melanoma stages.

Human Melanoma-Associated Mast Cells Display a Distinct Transcriptional Signature Characterized by an Upregulation of the Complement Component 3 That Correlates With Poor Prognosis

“…In addition, we found that conditioned MCs could be a source of the pro-tumorigenic cytokine IL-8 ( 53 ); CCL2, a potent chemokine that recruits MDSCs and TAMs ( 54 , 55 ); IL-33 that modulates melanoma cell proliferation, angiogenesis, and regulatory T cells ( 11 , 56 , 57 ); and the pro-angiogenic cytokine IL-1β ( 58 , 59 ). Melanoma cells apart from TGF-β produce many cytokines, among them growth factors such as TGFA, EGF, and HGF as well as cytokines such as IL-6, IL-8, and IL-1β ( 60 – 63 ). All these factors could potentially induce the expression of IL-8, CCL2, and IL-1β in MCs.…”

BRAF Mutations in Melanoma: Biological Aspects, Therapeutic Implications, and Circulating Biomarkers

“…Montoyo-Pujol and colleagues analysed a large number of soluble factors in both supernatant and sera from patients: interestingly, they detected the same molecules except for CXCL1 (present only in the supernatant). This suggests a local production and PDGF-BB (only present in sera) acting as a chemotactic molecule and showing a specific role in lymphangiogenesis promoting tumour progression and metastasis [ 76 ]. Data obtained by the authors indicated that PDGF-BB is not produced by melanoma cells, suggesting a possible secretion as systemic feedback to melanoma development.…”

Melanoma Tumour Vascularization and Tissue-Resident Endothelial Progenitor Cells

“…Tumour-derived exosomes released by melanoma cells are loaded by pro-angiogenic factors, such as VEGFA and MMPs. Serum analysis of human melanoma cells demonstrated the high expression of VEGFA, which could be utilised to monitor different melanoma stages [ 34 ]. Moreover, VEGFA expression is correlated with the progression of cutaneous melanoma [ 35 ].…”