Melanoma, while less prevalent than other skin cancers, remains the most lethal and aggressive type, posing significant treatment challenges. Photodynamic therapy (PDT) offers a promising, less invasive alternative to conventional therapies. In this study, we explored the potential of methylene violet 3RAX (MV), a phenazine-family photosensitizer, for PDT applications through in vitro assays and Langmuir monolayer studies, focusing on its interactions with cell lipid extract membranes derived from two melanoma lineages, A375 and SH-4. Our results demonstrate that MV is non-cytotoxic in the absence of light irradiation but exhibits concentration-dependent cytotoxicity upon photoactivation. Flow cytometry confirmed late apoptosis as the dominant cell death pathway under irradiation. Langmuir isotherms revealed that MV adsorbs onto anionic head groups of the lipid monolayers, particularly interacting with phosphate groups, promoting molecular organization. Upon irradiation, significant material loss to the subphase was observed, suggesting photooxidative interactions with lipid tail unsaturations, leading to hydroperoxidation, chain cleavage, and membrane destabilization. These findings highlight MV dual role as an effective photosensitizer and a molecular probe for membrane interactions, providing new insights into its mechanisms of action in PDT.
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