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目的:研究穿膜肽(CPPs)对载三氧化二砷蛋白微球进入细胞内的促进作用。方法:通过交联固化的方法制备三氧化二砷蛋白微球,N-羟基琥珀酰亚胺基-3-(2-吡啶基二硫)-丙酸脂(SPDP)交联的方法制备穿膜肽-增强型绿色荧光蛋白-三氧化二砷蛋白微球偶联物(As2O3-NS-CPPs-EGFP)。采用还原电泳、荧光显微镜和扫描电镜观察的方法鉴定As2O3-NS-CPPs-EGFP偶联物,激光共聚焦显微镜和透射电镜观察穿膜肽对蛋白微球进入细胞的促进作用,细胞内砷浓度测定明确药物细胞内传送效率。结果:CPPs-EGFP与三氧化二砷蛋白微球之间通过共价健连接,与穿膜肽偶联的三氧化二砷蛋白微球进入细胞内速度明显要快于单纯的三氧化二砷蛋白微球,用As2O3-NS-CPPs-EGFP处理组膀胱肿瘤细胞内砷浓度明显高于As2O3-NSEGFP处理组。结论:CPPs-EGFP具有促进As2O3-NS进入膀胱肿瘤细胞内的作用。
Objective: To study the promoting effect of penetrating peptide (CPPs) on the intracellular loading of arsenic trioxide-loaded protein microspheres. METHODS: Crosslinking and curing methods were used to prepare arsenic trioxide protein microspheres and crosslinked with N-hydroxysuccinimidyl-3- (2-pyridyldithio) propionate (SPDP) Type green fluorescent protein-arsenic oxide protein microsphere conjugate (As2O3-NS-CPPs-EGFP). The As2O3-NS-CPPs-EGFP conjugate was identified by reduction electrophoresis, fluorescence microscopy and scanning electron microscopy. The effects of penetrating peptide on the entry of protein microspheres into cells were observed by laser confocal microscopy and transmission electron microscopy. The intracellular arsenite concentration Clear drug intracellular delivery efficiency. Results: CPPs-EGFP and arsenic oxide arsenic protein microspheres were covalently linked to each other, and the rate of penetration into the cells with arsenic trioxide-loaded protein microspheres coupled with penetrating peptide was significantly faster than that of pure arsenic trioxide-containing protein microspheres. As2O3-NS-CPPs The arsenic concentration in bladder cancer cells inEGFP-treated group was significantly higher than that in As2O3-NSEGFP-treated group. Conclusion: CPPs-EGFP can promote As2O3-NS into bladder tumor cells.