Ozone therapy is an effective medical treatment for different diseases like mucositis, psoriasis, acute pain, neurovascular diseases and cancer. Emerging evidence indicates that ozone, a strong oxidant, could effectively improve organ ischemia-reperfusion, herniated disks and skin ulcers in clinical model with interesting anti-inflammatory properties through inhibition of NF-κB activation in acute and chronic disease. The aim of this study is based on the study of the biological effects of ozone in human melanoma cancer cells in order to investigate about its possible use in association to common therapy. Specifically, human melanoma cells were pre exposed or not to pro-inflammatory condition (Lipopolysaccharides) and administration of ozone at different concentration was performed in order to evaluate different biological parameters; cell viability, Measurement of Mitochondrial Matrix Potential (MMP), Evaluation of p65-Nfkb and changes in secretion of interleukins and growth factors involved in melanoma growth, survival and chemo-resistance: IL-1, IL-8, IL-6, TNF-α, IL-9, TGF-β, IL-19, VEGF, MMP-2, MMP-9 and IL-17 by ELISA methods. Results obtained shown ability of ozone to decrease cell viability up to 75% compared to control after 24h of incubation and inhibit all interleukins analyzed and involved in melanoma cell survival and drug resistance. Ozone at 50 ?g/ml also decrease of 60% the activation of the pro-inflammatory mediator p65Nfkb and inhibit the Nitric oxide production under pro-inflammatory condition. However, taking the several limitations of this study, further biological preclinical investigations are being carried out in order to understand the potential of ozone as possible adjuvant agent in therapy of melanoma.