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AIM:To prepare thymidine kinase gene (TK gene) nanoparticlesand to investigate the expression of TK gene.METHODS:Poly(D,L-lactic-co-glycolic acid) (PLGA),abiodegradable and biocompatible polymer,was used toprepare recombinant plasmid p~(EGFP-AFP) nanoparticles by adouble-emulsion evaporation technique.Characteristics ofthe nanoparticles were investigated in this study,includingmorphology,entrapment efficiency,and tissue distribution.The expression of TK gene was also investigated by MTTassay,by which the viable cells were determined after theaddition of ganciclovir (GCV).The enhanced green fluorescentprotein (EGFP) expression in human hepatocellular carcinomaSMMC-7721 cells and normal parenchymal Chang liver cellswere assessed by flow cytometry.RESULTS:The prepared plasmid-nanoparticles had regularspherical surface and narrow particle size span with a meandiameter of 72±12 nm.The mean entrapment efficiencywas 91.25%.A total of 80.14% DNA was found to belocalized in the livers after 1-h injection with ~(32)P-DNA-PLGAnanoparticles in mouse caudal vein.The expression of DNAencapsulated in nanoparticles was much higher than that innaked DNA,and human hepatocellular carcinoma SMMC-7721 cells were more sensitive to GCV than human normalparenchymal Chang liver ceils.CONCLUSION:The enhanced transfection efficiency andstronger ability to protect plasmid DNA from being degradedby nucleases are due to nanoparticles encapsulation.
AIM: To prepare thymidine kinase gene (TK gene) nanoparticles and to investigate the expression of TK gene. METHODS: Poly (D, L-lactic-co- glycolic acid) (PLGA), abiodegradable and biocompatible polymer, was used toprepare recombinant plasmid p ~ (EGFP-AFP) nanoparticles by adouble-emulsion evaporation technique. Characteristics of the nanoparticles were investigated in this study, including morphology, entrapment efficiency, and tissue distribution. The expression of TK gene was also investigated by MT Assay, by which the viable cells were determined after the addition of ganciclovir (GCV) .The enhanced green fluorescent protein (EGFP) expression in human hepatocellular carcinomaSMMC-7721 cells and normal parenchymal Chang liver cellswere assessed by flow cytometry.RESULTS: The prepared plasmid-nanoparticles had regularspherical surface and narrow particle size span with a meandiameter of 72 ± 12 nm. The mean entrapment efficiency was 91.25%. A total of 80.14% DNA was found to belocalized in the livers after 1- h injection with ~ (32) P-DNA-PLGAnanoparticles in mouse caudal vein. The expression of DNA was encapsulated in nanoparticles was much higher than that innaked DNA, and human hepatocellular carcinoma SMMC-7721 cells were more sensitive to GCV than human normal parenchymal Chang liver ceils .CONCLUSION: The enhanced transfection efficiency and enhanced ability to protect plasmid DNA from being degraded by nucleases due due to nanoparticles encapsulation.