CD133^+细胞的干性鉴定及^(131)Ⅰ-CD133抗体对其体内外抑制作用  

Identification of the stem property of CD133^+liver cancer cells and inhibition effect of ^(131)Ⅰ-labeled anti-CD133 mAb on them in vitro and vivo

作　　者：段丽群[1] 侯妍利[1] 陈兴月[1] 唐敏[1] 康强强[1] 舒锦[1] 李少林[1] 

机构地区：[1]重庆医科大学基础医学院放射医学教研室,重庆400016

出　　处：《重庆医科大学学报》2014年第7期932-937,共6页Journal of Chongqing Medical University

基　　金：国家自然科学基金面上资助项目(编号:30970843、81171365)

摘　　要：目的:研究CD133+细胞的干性鉴定和131I-CD133抗体在体内外对人肝癌CD133+-HepG2干细胞的抑制作用。方法:氯胺T法制备并鉴定131I-CD133抗体;免疫磁珠(magnetic-activated cell sorting,MACS)分选CD133+-HepG2细胞;流式细胞仪(flow cytometry,FCM)检测分选前后CD133表达率;体外克隆形成实验、成球实验和体内成瘤实验验证其干细胞特性;将分选出的CD133+细胞分组为CD133抗体、131I、131I-CD133抗体和131I+CD133抗体4个组,MTT法检测不同处理后不同组中CD133+细胞生长抑制率;成功构建人肝癌CD133+-HepG2移植瘤模型;随机分4组,1次/2 d给予尾静脉用药,共14次。4周后,处死小鼠,比较肿瘤的体积、质量、计算抑瘤率;HE染色观察肿瘤组织病理学改变。结果:131I-CD133抗体标记率为89.34%,放化纯度为98.21%。流式显示分选前后CD133表达率分别为(1.78±0.54)%和(98.46±0.97)%。成球实验、克隆形成实验和裸鼠成瘤实验显现CD133+细胞相对于CD133-细胞更具有干细胞特性。131I-CD133抗体治疗组体外对细胞抑制率及体内抑瘤率明显高于其余各实验组,差异具有统计学意义(P<0.05)。结论:131I-CD133抗体在体内外均能有效抑制人肝癌CD133+-HepG2细胞的生长。Objective:To study the identification the stem property of CD133+ liver cancer cells and inhibition effect of 131I-labeled monoclonal antibody(mAb)against CD133 on them in vitro and vivo. Methods:The monoclonal antibody CD133 labeled with 131I was prepared using the chloramines-T method and its stability was evaluated. Magnetic-activated cell sorting(MACS)was used to isolate CD133+and CD133-cells from HepG2 cells. Flow cytometry(FCM)was used to detect the expression of CD133 before and after the cell isolation. The stem cell properties of sorted CD133+cells were validated by sphere-forming assay,colony formation assay in vit ro and tumorigenesis experiment in vivo. There were four groups including131I-labeled anti-CD133 mAb group,131I group,CD133 mAb group and131I+CD133 mAb group. The inhibitory effects of different treatments on the proliferation of CD133+cells were measured by MTT assy. The animal model was established by subcutaneous inoculation of CD133+-HepG2 cells(5×104)to right front legs of BALB/c mice. After tumor model being established,12 mice were randomly divided into 4 groups. Drugs were injected into the tail vein of the nude mice at a frequency of one time/2 d(14 times in total). The tumor size and volume were measured twice a week after the treatment and the tumor inhibitory rate was calculated. After 4 weeks,the mice were sacrificed for pathological examination. Results:The labeling ratio of the131I-labeled anti-CD133 mAb was 89.34% and the radiochemical purity was 98.21%. The sorted CD133+cells showed a high purity(98.46±0.97)% compared with that before cell sorting(1.78±0.54)%. CD133+ cells showed a high tumor sphere formation ability and turmorigenesis capacity compared with those of CD133-cells. The tumor inhibitory rate of CD133+cells was significantly higher in131I-labeled anti-CD133 mAb group than in131I group,CD133 mAb group and blank control group in vivo and vitro(P<0.05). Conclusion:131I-labeled anti-CD133 mAb can effectively inhibit the growth of CD133+-HepG2 cells in vivo and vit

关 键 词：CD133 肝癌干细胞 131I 单克隆抗体 裸鼠 

分 类 号：R735.7[医药卫生—肿瘤]