In recent years

nanosecond pulsed electric fields (nsPEFs) have garnered significant attention as a potential cancer treatment method. This study aims to investigate the therapeutic effects of nsPEFs on melanoma and explores their biological effects in inducing pyroptosis in melanoma cells and stimulating the body's immune response. The B16F10 mouse melanoma cell line was chosen as the experimental subject. The cells were treated with nsPEFs (200 ns

1 Hz) both in vitro and in vivo

and cytological

molecular biological

and immunological techniques were used to assess cell viability

pyroptosis

expression levels of related proteins and molecules

and immune-related molecules.The experimental results showed that nsPEFs stimulation caused bubbling on the cell membrane surface and increased membrane permeability. The expression of cleaved Caspase-1 and cleaved GSDMD proteins increased

indicating that nsPEFs effectively induced pyroptosis in melanoma cells. The expression of damage associated molecular patterns (DAMPs) such as calreticulin (CRT)

heat shock protein 70 (HSP70)

and high mobility group protein B1 (HMGB1) was also elevated

suggesting that nsPEFs-induced cell death is immunogenic.Following nsPEFs stimulation

tumor volume gradually diminished

and a large number of activated dendritic cells (DCs) and CD8+ T cells appeared in the tumor tissue. Serum levels of IL-12

TNF-α

and IFN-γ increased

while levels of IL-10 and TGF-β1 decreased

suggesting that mice might initiate immune response pathways through the release of these factors

participating in the anti-tumor effect after nsPEFs ablation. Tumor challenge experiments revealed that

under the accompaniment of specific immune responses

tumor growth in the nsPEFs-stimulated group was significantly inhibited. The results indicate that nsPEFs have anti melanoma effects and can short-termly stimulate the body's immune system to enhance treatment efficacy.