Aim To survey the uptake behavior and subcellular distribution of antisense oligodeoxynucleotide polymethacrylate submicroparticles （AS-ODN-SMP） and infer its mechanism in MGC cell lines. Methods MGC cells were incu...Aim To survey the uptake behavior and subcellular distribution of antisense oligodeoxynucleotide polymethacrylate submicroparticles （AS-ODN-SMP） and infer its mechanism in MGC cell lines. Methods MGC cells were incubated at certain concentration of AS-ODN-SMP or AS-ODN for 8 h at 4 ℃ or 37 ℃. Then the fluorescence oligodeoxynucleotide- labeled cells were counted by flow cytometer and the intracellular fluorescence intensity was determined after incubated with chloroquine for 2 h. Results Cellular uptake of oligodeoxynucleotides was significantly increased following application of AS-ODN-SMP and total intracellular fluorescence intensity was enhanced by 683 folds with the vehicle concentration of 20 μg·mL-1. AS-ODN-SMP entranced to cells profoundly with temperaturedependent manner. Rare cells took on fluorescence when incubated at 4 ℃, while 37 ℃ they were significantly increased. But the intracellular fluorescence intensity appeared same level in present or absent of chloroquine. Conclusion With the help of polyacrylate submicroparticles, oligonucleotides efficiently entranced the cells via endocytosis and could successfully escape the degradation in lysosome.展开更多
Aim To construct an efficient recombinant viral vector for gene therapy.Methods(First-generation) adenovirus(Ad) vector was modified with the RGD peptide inserted into the fiber.Both in vitro and in vivo experiments o...Aim To construct an efficient recombinant viral vector for gene therapy.Methods(First-generation) adenovirus(Ad) vector was modified with the RGD peptide inserted into the fiber.Both in vitro and in vivo experiments of gene expression in different tumor cells with conventional and recombinant vectors were conducted.RT-PCR was used for detecting the expression of coxackievirus and adenovirus receptor and integrin at the surface of Meth-A cells.Results Fiber-mutant adenovirus vector showed a notably enhanced gene expression in A2058,B16BL6,OV-HM,and Meth-A tumor cells compared with that of conventional ones.In vivo study carried out using Meth-A tumor-bearing mice also demonstrated that the intra-tumoral injection of recombinant adenovirus induced strong gene expression in these CAR-deficient tumor cells. Conclusion The recombinant vector can be a promising one for effective cancer gene therapy.展开更多
文摘Aim To survey the uptake behavior and subcellular distribution of antisense oligodeoxynucleotide polymethacrylate submicroparticles （AS-ODN-SMP） and infer its mechanism in MGC cell lines. Methods MGC cells were incubated at certain concentration of AS-ODN-SMP or AS-ODN for 8 h at 4 ℃ or 37 ℃. Then the fluorescence oligodeoxynucleotide- labeled cells were counted by flow cytometer and the intracellular fluorescence intensity was determined after incubated with chloroquine for 2 h. Results Cellular uptake of oligodeoxynucleotides was significantly increased following application of AS-ODN-SMP and total intracellular fluorescence intensity was enhanced by 683 folds with the vehicle concentration of 20 μg·mL-1. AS-ODN-SMP entranced to cells profoundly with temperaturedependent manner. Rare cells took on fluorescence when incubated at 4 ℃, while 37 ℃ they were significantly increased. But the intracellular fluorescence intensity appeared same level in present or absent of chloroquine. Conclusion With the help of polyacrylate submicroparticles, oligonucleotides efficiently entranced the cells via endocytosis and could successfully escape the degradation in lysosome.
文摘Aim To construct an efficient recombinant viral vector for gene therapy.Methods(First-generation) adenovirus(Ad) vector was modified with the RGD peptide inserted into the fiber.Both in vitro and in vivo experiments of gene expression in different tumor cells with conventional and recombinant vectors were conducted.RT-PCR was used for detecting the expression of coxackievirus and adenovirus receptor and integrin at the surface of Meth-A cells.Results Fiber-mutant adenovirus vector showed a notably enhanced gene expression in A2058,B16BL6,OV-HM,and Meth-A tumor cells compared with that of conventional ones.In vivo study carried out using Meth-A tumor-bearing mice also demonstrated that the intra-tumoral injection of recombinant adenovirus induced strong gene expression in these CAR-deficient tumor cells. Conclusion The recombinant vector can be a promising one for effective cancer gene therapy.
