基于靶向低密度脂蛋白受体蛋白1的载药嵌段共聚物抗胶质瘤作用的体外实验研究

Application of drug-loaded block copolymers targeting LRP1 for treatment of glioma cells in vitro

目的研究靶向低密度脂蛋白受体蛋白1(LRP1)的载药嵌段共聚物(ANPs/CPT)对胶质瘤细胞的增殖抑制和促进凋亡的作用。方法采用Western blotting技术检测LRP1在大鼠胶质瘤细胞(C6细胞)和脑毛细血管内皮细胞(BCECs)的表达水平。用激光共聚焦显微镜测定C6细胞和BCECs对Angiopep-2修饰的靶向未载药嵌段共聚物(ANPs)和非靶向对照NPs的摄取量;采用噻唑蓝(MTT)比色法检测ANPs/CPT抑制C6细胞和BCECs增殖的作用;采用流式细胞仪分析不同CPT终浓度(10μg/mL,20μg/mL)的ANPs/CPT促C6细胞凋亡作用。结果LRP1在C6和BCECs细胞膜表面高表达。C6细胞对ANPs的摄取量显著高于NPs(P<0.01);ANPs/CPT对C6细胞的增殖抑制作用显著高于NPs/CPT(P<0.05),但对BCECs的增殖抑制作用与NPs/CPT无显著差异(P>0.05)。CPT终浓度10μg/mL、20μg/mL的ANPs/CPT与NPs/CPT作用于C6细胞48 h后的凋亡率分别为(16.1±0.9)%、(30.4±1.1)%和(18.8±0.9)%、(40.3±1.2)%,差异均有统计学意义(均P<0.05)。结论ANPs/CPT有显著的靶向抑制胶质瘤细胞增殖及促进其凋亡的作用,具有潜在的治疗胶质瘤的应用价值。

Objective To study the effects of drug loaded block copolymers(ANPs/CPT)targeting low density lipoprotein receptor protein 1(LRP1)on the proliferation and apoptosis of glioma cells.Methods The expression levels of LRP1 in rat glioma cells(C6 cells)and brain capillary endothelial cells(BCECs)were detected by Western blotting.The uptake of angiope2 modified targeted drug free block copolymers(ANPs)and non targeted control NPs by C6 cells and BCECs were measured by laser confocal microscopy.The inhibitory effect of ANPs/CPT on the proliferation of C6 cells and BCECs was detected by thiazole blue(MTT)colorimetry.Different CPT final concentrations were analyzed by flow cytometry(10μg/mL,20μg/mL)of ANPs/CPT promoted C6 cell apoptosis.Results LRP1 was highly expressed on the surface of C6 and BCECs cell membrane.The uptake of ANPs by C6 cells was significantly higher than that of NPs(P<0.01).The inhibitory effect of ANPs/CPT on the proliferation of C6 cells was significantly higher than that of NPs/CPT(P<0.05),but there was no significant difference between ANPs/CPT and NPs/CPT(P>0.05).The apoptosis rates of C6 cells treated with ANPs/CPT and NPs/CPT at the final concentration 10μg/mL,20μg/mL of CPT for 48 hours were(16.1±0.9)%,(30.4±1.1)%and(18.8±0.9)%,(40.3±1.2)%.The differences were statistically significant(all P<0.05).Conclusion ANPs/CPT can significantly inhibit the proliferation and promote the apoptosis of glioma cells,and has potential application value in the treatment of glioma.