TRANSFECTION OF GFP PLASMIDS BY LIPOSOMAL CARRIERS IN B16 MELANOMA CELLS AND MELANOMA MODELS IN VIVO

Abstract

Abstract. Background: Penetration of plasmid DNA into the nucleus by a passive process is ineffective due to the barrier effect of lipid membranes and diffusion barriers inside the cell. An effective gene delivery system should ensure binding to the target cell, penetration by endocytosis, protection from degradation and, ultimately, delivery of the expression plasmid to the nucleus. The distribution routes of transfected agents play a crucial role in achieving the targeted effect of drugs. In our experiment, plasmid DNA of the reporter protein GFP was used to study the distribution pathways. Aim: to evaluate the distribution of GFP reporter protein in B16 melanoma cells and distant organs of C57Bl6 mice as a method for determining the effectiveness of transfection of genetically engineered structures for the diagnosis and therapy of tumors in vivo. Materials and methods: An in vitro study was conducted on skin melanoma cell lines B16. Cells were transfected with a plasmid of the reporter protein GFP. In vivo, the study was conducted on mature female mice of the C57Bl/6 line, to which tumor cells were transplanted and GFP plasmid was intraperitoneally injected using commercial transfectants Lipofectamine 3.0 and Invivofectamine. The efficiency of transfection in organ and tumor cells was evaluated using the cell imaging station Floid. Results: A high survival rate of B16 melanoma cells was established and GFP+ B16 melanoma cells were obtained within 48 hours. We found that with intraperitoneal administration of a lipocomplex containing GFP plasmid, both with the use of Lipofectamine 3.0 transfectant and Invivofectamine, effective incorporation into liver and kidney parenchyma cells occurs. The penetration and retention of GFP reporter protein in liver, kidney and tumor node cells was revealed. Conclusion: The effectiveness of transfection of GFP plasmid into liver, kidney, and melanoma B16 tumor node cells of C57Bl6 mice has been proven. The data obtained indicate the prospects for the use of transfecting agents for penetration into target cells of genetically engineered structures for the purpose of diagnosis and therapy of tumors in vivo.