Combined cold atmospheric plasma and vemurafenib suppress MITF–P۲۱ signaling and MMP‑driven metastasis in cutaneous melanoma

Objective(s): This study investigated the anti-tumor efficacy of combined cold atmospheric plasma (CAP) and Vemurafenib (VEM) treatment (CAP plus VEM) in cutaneous melanoma, focusing on the MITF–P۲۱ axis and MMP‑driven metastasis.Materials and Methods: The therapeutic effects of CAP and VEM were evaluated using in vitro and in vivo models. B۱۶‑F۱۰ melanoma and L۹۲۹ fibroblast cell lines were used to assess long‑term cell proliferation and migration using clonogenic and wound‑healing assays. In vivo, melanoma tumors were induced in C۵۷BL/۶ mice, followed by treatment with CAP, VEM, or CAP plus VEM. Tumor tissues and major organs were collected for histopathological evaluation and molecular analysis. Gene expression levels of MITF, P۲۱, MMP‑۲, and MMP‑۹ were quantified by qRT‑PCR in both experimental settings.Results: CAP and VEM significantly inhibited melanoma cell proliferation and migration, with the CAP plus VEM showing the most pronounced effect. In vitro, all treatments markedly down-regulated MITF, P۲۱, and MMP-۲ gene expression (P<۰.۰۰۰۱). Histopathological analysis revealed extensive tumor necrosis, reduced cellularity, and improved tissue architecture in the CAP plus VEM group, accompanied by fewer metastatic alterations in major organs. qRT‑PCR of tumor tissues confirmed significant down-regulation of MMP‑۲ and MMP‑۹ after CAP, VEM, and especially the CAP plus VEM group (P<۰.۰۰۰۱). The modest in vivo changes in MITF and P۲۱ may reflect microenvironmental influences.Conclusion: The CAP plus VEM enhances anti-tumor efficacy and suppresses metastasis‑related pathways in melanoma, both in vitro and in vivo. This suggests a promising complementary therapeutic strategy.

کلیدواژه ها:

Animal ، In vitro ، In vivo ، Melanoma ، Matrix metalloproteinases Models ، Plasma gases ، Vemurafenib