反义寡核苷酸聚甲基丙烯酸酯亚微粒入胞行为的研究  

Cellular uptake behavior of antisense oligodeoxynucleotides polymethacrylate submicroparticles

作　　者：王文喜[1] 梁文权[2] 宋必卫[1] 陈海靓[2] 

机构地区：[1]浙江工业大学药学院,浙江杭州310014 [2]浙江大学药学院,浙江杭州310031

出　　处：《药学学报》2005年第10期950-953,共4页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(30371692)

摘　　要：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.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 temperature-dependent 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.

关 键 词：聚甲基丙烯酸酯 砸微粒 反义寡核苷酸 入胞行为 

分 类 号：R943[医药卫生—药剂学]