转铁蛋白受体单克隆抗体纳米载药系统治疗白血病的基础研究

Basic Research on Therapy of Leukemia with Nanoparticles Drug Delivery System Modified by Transferrin Receptor Monoclonal Antibody

目的:探讨转铁蛋白受体单克隆抗体(TfR mAb)纳米载药系统靶向治疗急性白血病的效果及其潜在的抗肿瘤机制。方法:合成纳米载药粒TfR mAb-聚乳酸-羟基乙酸(PLGA)-聚L-赖氨酸(PLL)-聚乙二醇(PEG)-柔红霉素(DNR)。急性髓性白血病细胞株HL60细胞经药物干预后,倒置荧光显微镜下观察细胞内DNR的累积;流式细胞技术(FCM)检测HL60细胞内DNR浓度及细胞凋亡率;Western blot测定凋亡相关蛋白Cleaved-caspase 3的表达量。结果:单药DNR组和TfR mAb-PLGA-PLL-PEG-DNR组HL60细胞内可见DNR自发荧光累积,且TfR mAb-PLGA-PLL-PEG-DNR组细胞内DNR浓度高于单药DNR组(P<0.05);FCM检测结果显示,TfR mAb-PLGA-PLL-PEG-DNR组细胞凋亡率高于单药DNR组(P<0.05);Western blot检测结果证实,TfR mAb-PLGA-PLL-PEG-DNR组凋亡相关蛋白Cleaved-caspase 3表达量明显高于单药DNR组(P<0.05)。结论:TfR mAb-PLGA-PLL-PEG纳米载药系统将化疗药物靶向作用于肿瘤细胞HL60,可通过凋亡途径增加药物的抗肿瘤能力。

Objective:To investigate the therapeutic effect of targeted drug-loaded nanoparticles modified by transferrin receptor monoclonal antibody(TfR mAb)on acute leukemia and its potential anti-tumor mechanism.Methods:Nanoparticles drug delivery system,which was composed of poly(lactic-co-glycolic acid),poly-l-lysine,polyethylene glycol,TfR mAb(TfR mAb-PLGA-PLL-PEG)-daunorubicin(DNR),was first synthesized.After drug intervention,the intracellular accumulation in leukemia HL60 cells was observed under a fluorescent microscope and concentration of DNR was determined by flow cytometry(FCM).Meanwhile,cell apoptosis rate was measured by FCM and the expression levels of apoptosis related protein Cleaved:-caspase 3 was determined by Western blot.Results:Under an inverted fluorescent microscope,intracellular accumulation of DNR autofluorescence in HL60 cells was observed in both TfR mAb-PLGA-PLL-PEG-DNR group and DNR group.FCM analysis showed that the intracellular concentration of DNR in TfR mAb-PLGA-PLL-PEG-DNR group was higher than that in DNR group(P<0.05).The apoptotic rate of HL60 cells in TfR mAb-PLGA-PLL-PEG-DNR group was higher than that of DNR group(P<0.05).Moreover,the expression levels of apoptosis related protein Cleaved-caspase 3 in TfR mAb-PLGA-PLL PEG-DNR group was significantly higher than that in DNR group(P<0.05).Conclusion:TfR mAb-PLGA-PLL-PEG nanoparticle drug delivery system can target chemotherapy drugs to leukemia cells and enhance anticancer ability through apoptotic pathway.