反义寡核苷酸聚甲基丙烯酸酯亚微粒入胞行为的研究反义寡核苷酸聚甲基丙烯酸酯亚微粒入胞行为的研究

反义寡核苷酸(antisense oligodeoxynucleotides,AS-ODN)是一种能与特定的DNA或RNA特异性结合并阻断基因表达的生物效应分子,具有作用强、特异性高等优点,受到广泛关注1]。然而由于AS-ODN为亲水性大分子,细胞通透性差,使得它的应用受到极大限制。为了增加AS-ODN在体内的稳定性

Cellular uptake behavior of antisense oligodeoxynucleotides polymethacrylate submicroparticles

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 degrees C or 37 degrees C. 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 microg x mL(-1). AS-ODN-SMP entranced to cells profoundly with temperature-dependent manner. Rare cells took on fluorescence when incubated at 4 degrees C, while 37 degrees C 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.