The role of vitamin C in melanoma cell death via activation of cytochrome C and TNF-α protein expression.

This study investigates the effect of vitamin C on cytochrome c and TNF-α expression, representing intrinsic and extrinsic apoptotic pathways in B16-F10 melanoma cells. B16-F10 melanoma cells (ATCC® CRL-6475™) were treated with vitamin C for 12 and 24 h. Expression of cytochrome c, TNF-α, caspase-3, and caspase-9 proteins was evaluated using in-cell western (ICW) assay. RT-qPCR was used to assess gene expression of Caspase-8, IL-6, PARP-γ, and TGF-β1. Morphological signs of cell death appeared within 12 h. A significant increase in cytochrome c expression at 3,000 µM concentrations of vitamin C was observed (p = 0.002), along with increases at 3,500 µM (p = 0.023), 4,000 µM (p = 0.0001), and 5,000 µM (p = 0.0001). Caspase-3 expression significantly increased at 3,000 µM (p = 0.012) and 5,000 µM (p = 0.048), while caspase-9 was not significant. TNF-α expression also increased significantly from 3,000 to 5,000 µM (all p < 0.01). Caspase-8 and PARP-γ mRNA levels were upregulated at 3,000 and 5,000 µM (p < 0.05). IL-6 and TGF-β1 changes further supported apoptosis activation. Vitamin C induces apoptosis in B16-F10 melanoma cells via intrinsic and extrinsic pathways, particularly at 3,000 and 5,000 µM concentrations, as shown by cytochrome c, TNF-α, and caspase-3 expression.