Copper nitroprusside analogue nanoparticles against melanoma: Detailed in vitro and in vivo investigation

Melanoma is the most invasive and lethal form of skin cancer that arises from the malignant transformation of specialized pigmented cell melanocytes. Nanomedicine represents an important prospect to mitigate the difficulties and to provide significant benefit to cure melanoma. In the present manuscript, we have investigated the in vitro and in vivo therapeutic efficacy of copper nitroprusside analogue nanoparticles (abbreviated as CuNPANP) towards melanoma. Initially, in vitro anti-cancer activities of CuNPANP towards melanoma cells (B16F10) are evaluated by several experiments such as [methyl 3H]-thymidine incorporation assay, cell cycle & apoptosis assays using FACS analysis, ROS generation using DCFDA, DHE and DAF2A reagent, internalization of nanoparticles through ICP-OES analysis, co-localization of the nanoparticles using confocal microscopy, JC-1 staining to investigate the mitochondrial membrane potential (MMP) and immunofluorescence studies to analyze the expressions of cytochrome-c, Ki-67, E-cadherin as well as phalloidin staining to analyze the cytoskeletal integrity. Further, the in vivo therapeutic effectiveness of the nanoparticles has been established towards malignant melanoma by inoculating B16F10 cells in the dorsal right abdomen of C57BL/6J mice. Intraperitoneal administrations of CuNPANP inhibit tumor growth and increase the survivability of melanoma mice. The in vivo immunofluorescence studies (Ki-67, CD-31, E-cadherin) and TUNEL assay further supports the anti-proliferative and apoptosis inducing potential of CuNPANP, respectively. Finally, the various signaling pathways and molecular mechanisms involved for anti-cancer activity are further evaluated by Western blot analysis. The results altogether offer the use of copper-based nanomedicine for the treatment of malignant melanoma.