三氧化二砷聚乙二醇-聚乳酸偶联人源抗血管内皮细胞生长因子受体2隐形纳米粒的抗肝癌机制探讨  

Anti- hepatoma Mechanism of Human Anti- VEGFR- 2 / As_2O_3- PEG- PLA Stealth Nanoparticles

作　　者：殷香保[1] 邬林泉[1] 黄跃英[1] 黄明文[1] 王恺[1] 周凡[1] 余新[1] 

机构地区：[1]南昌大学第二附属医院肝胆外科,江西省南昌市330006

出　　处：《中国全科医学》2015年第23期2805-2809,共5页Chinese General Practice

基　　金：国家自然科学基金资助项目(81060187);江西省自然科学基金资助项目(2008GQY0050);江西省卫生厅科技计划项目(20131078)

摘　　要：目的探讨以聚乙二醇-聚乳酸(PEG-PLA)为载体材料包载的三氧化二砷(As2O3),同时偶联人源抗血管内皮细胞生长因子受体2(VEGFR-2)的隐形纳米粒的抗肝癌机制。方法以PEG-PLA为载体材料,W/O/W型超声乳化制备As2O3纳米粒,同时偶联具有肝癌靶向作用的VEGFR-2,得到人源抗VEGFR-2/As2O3-PEG-PLA纳米粒,同时以相同方法制作香豆素6(C6)-PEG-PLA及抗VEGFR-2/C6-PEG-PLA纳米粒。测定纳米粒粒径分布、Zata电位;MTT法计算As2O3、As2O3-PEG-PLA及抗VEGFR-2/As2O3-PEG-PLA对Bel 7402肝癌细胞IC50值;采用流式细胞实验和激光共聚焦显微镜实验检测人肝癌Bel 7402细胞摄取C6-PEG-PLA及抗VEGFR-2/C6-PEG-PLA纳米粒的速度和强度;将18只肝癌裸鼠采用随机数字表法分为3组,每组分别经尾静脉注射As2O3、As2O3-PEG-PLA、抗VEGFR-2/As2O3-PEG-PLA溶液,给药后12 h处死小鼠,取肿瘤、脾、心、肺、肝、肾等组织,测试其中As2O3浓度;再将另24只肝癌裸鼠采用随机数字表法分为4组,每组分别经尾静脉注射As2O3、As2O3-PEG-PLA、抗VEGFR-2/As2O3-PEG-PLA及0.9%氯化钠溶液,给药后第17 d,处死小鼠,计算肿瘤抑制率。结果抗VEGFR-2/As2O3-PEG-PLA平均粒径为(141.9±13.2)nm(PDI=0.23),呈正态分布,Zata电位为(10.2±1.1)m V。As2O3、As2O3-PEG-PLA、抗VEGFR-2/As2O3-PEG-PLA的IC50值分别为(1.53±0.22)μmol/L,(2.30±0.18)μmol/L和(1.80±0.22)μmol/L。不同剂型IC50值比较,差异有统计学意义(F=4.503,P=0.018)。不同时间点人肝癌Bel 7402细胞摄取抗VEGFR-2/C6-PEG-PLA的速度均高于C6-PEG-PLA,差异有统计学意义(t=2.012,P=0.045;t=2.231,P=0.035;t=2.311,P=0.022)。As2O3组不同组织As2O3含量比较,差异有统计学意义(F=5.568,P=0.018);As2O3-PEG-PLA组不同组织As2O3含量比较,差异有统计学意义(F=4.659,P=0.034);抗VEGFR-2/As2O3-PEG-PLA组不同组织As2O3含量比较,差异有统计学意义(F=5.001,P=0.022)。对照组、As2O3组、As2O3-PEG-PLA组和抗VEGFR-2/As2O3-PEG-PLA组肿瘤抑制率分别为32.6%�Objective To investigate the anti -hepatoma mechanism of human anti -VEGFR-2/As2 O3 -PEG-PLA stealth nanoparticles . Methods PEG-PLA was used as the vector material to prepare As 2 O3 nanoparticles with W/O/W ultrasonic emulsification method , and VEGFR-2 that is targeted at liver cancer was coupled to obtain human anti -VEGFR-2/As2 O3 -PEG-PLA nanoparticles.C6 -PEG-PLA and anti -VEGFR-2/C6 -PEG-PLA stealth nanoparticles were also prepared using the same method .Size distribution and Zata potential of the nanoparticles were characterized using laser particle＆amp;nbsp;analyzer; IC50 values of As2 O3 , As2 O3 -PEG-PLA and anti -VEGFR-2/As2 O3 -PEG-PLA against liver cancer cell Bel 7402 were determined using MTT method; laser scanning confocal microscope and flow cytometry were used to measure the speed and intensity of the uptake of C 6 -PEG-PLA and anti -VEGFR-2/C6 -PEG-PLA nanoparticles by liver cancer cell Bel 7402; using random number table method , we divided 18 nude mice with liver cancer into three groups and assigned the three groups to receive the injection of As 2 O3 , As2 O3 -PEG-PLA and anti -VEGFR-2/As2 O3 -PEG-PLA solutions respectively through caudal veins , and we killed the mice 12 months after the injection and obtained their tumors , spleens, hearts, lungs, livers and kidneys to measure the As 2 O3 concentration within these tissues; using random number table method , we divided another 24 nude mice with lung cancer and assigned the three groups to receive the injection of As 2 O3 , As2 O3 -PEG-PLA, anti-VEGFR-2/As2 O3 -PEG-PLA and 0.9% sodium chloride solutions respectively through caudal veins , and we killed the mice 17 days after the injection and calculated the tumor inhibiting rates . Results The average particle size of anti -VEGFR-2/As2O3 -PEG-PLA was （141.9 ±13.2） nm （PDI=0.23） with a normal distribution.The Zata potential of anti-VEGFR-2/As2 O3 -PEG-PLA was （ 10.2 ±1.1 ） mV.The IC50 values of As2 O3 , As2 O3 -PEG-PLA and anti -VEGFR-2/As2 O3 -PEG-PLA were （ 1.53 ±0.22

关 键 词：砷剂 聚乙二醇-聚乳酸 血管内皮生长因子类 肝肿瘤 机制 

分 类 号：R735.7[医药卫生—肿瘤]