Zahra Yazdani
, Farshad Sohbatzadeh, Saeid Abediankenari, Monireh Golpour, Sadegh Fattahi, Alireza Rafiei
*
Abstract
Purpose: This study investigated the combined effects of cold atmospheric plasma (CAP) and vemurafenib (VEM) on malignant melanoma, focusing on redox signaling, apoptosis, and autophagy. Methods: Murine melanoma B16‑F10 cells and normal L929 fibroblasts were treated with CAP, VEM, or their combination. Cell viability was assessed using the MTT assay. Reactive oxygen and nitrogen species (RONS) and lipid peroxidation were measured to evaluate oxidative stress. Cell cycle distribution and apoptosis were analyzed by propidium iodide (PI) staining and Annexin V-FITC/PI flow cytometry. An in vivo B16‑F10 tumor‑bearing mouse model was utilized to evaluate tumor growth and molecular changes in tumor tissues. The expression of apoptosis‑ and autophagy‑related genes (BAX, BCL‑2, CASP3, LC3, and ATG5) was quantified by qRT‑PCR in vitro and in vivo experiments. Results: The combination treatment significantly reduced the viability of B16‑F10 melanoma cells with minimal effects on L929 fibroblasts. This reduction was associated with increased RONS production and lipid peroxidation. Cell cycle analysis indicated S‑phase arrest in melanoma cells, and apoptosis assays showed enhanced apoptotic cell death after the combination treatment. Molecular analysis revealed increased expression of BAX and CASP3, whereas BCL‑2 expression showed a decreasing trend. The treatment also increased autophagy‑related gene expression. In the mouse melanoma model, the combined therapy suppressed tumor growth and increased the expression of apoptosis‑ and autophagy‑related genes, including BAX, CASP3, ATG5, and LC3. Conclusion: CAP enhances the antitumor effects of VEM by promoting oxidative stress and activating apoptosis‑ and autophagy‑related gene expression. This combined approach may represent a promising strategy for melanoma treatment, although further studies are needed to confirm these findings.