Construction of a Golgi protein 73 modified liposome that mediates HSVtk induced liver cancer cell apoptosis

Abstract

Objective To construct a non-viral gene therapy vector (GP73 modified lipoplexes) and to assess its delivery efficiency and impact on apoptosis in hepatoma cell line HepG2 and normal liver cell line HL-7702. Methods Lipoplexes were conjugated with human GP73 using water-soluble carbodidimide, a crossing agent. Then, GP73-lipoplexes were used to carry pGenesil-l plasmid containing EGFP gene and transfect hepatoma cell line HepG2 and normal liver cell line HL-7702. Fluorescence microscopy and flow cytometry were used to assess the transfection efficiency. GP73-lipoplexes were then used to carry PBI-SUR-TK plasmid containing the therapeutic gene HSVtk and transfect hepatoma cells and normal hepatocytes. RT-PCR and Western blot were used to detect the expression of HSVtk in cells, and flow cytometry and CCK-8 assay were used to detect apoptosis. Results Compared with HL-7702 cells, HepG2 cells had higher transfection efficiency [(9.68±0.57)% vs (26.37±0.76)%], obvious HSVtk gene production, and significant apoptosis. Conclusion GP73 modified lipoplexes can be an effective tool to improve the transfection efficiency and therapeutic effects of exogenous genes in hepatoma cells. Key words: GP73; Lipoplexes; Hepatoma cell; Transfection efficiency; Apoptosis