靶向血管内皮生长因子受体2超声造影剂的制备及其体外寻靶能力实验研究  

Preparation of vascular endothelial growth factor receptor 2-targeted ultrasonic contrast agent and its ability of targeting tumor cells in vitro

作　　者：袁海霞[1] 汤阳[1] 汪瀚韬 孔文韬 王文平[1] 

机构地区：[1]复旦大学附属中山医院超声科,上海200032 [2]江苏省南京鼓楼医院超声科,江苏南京210008

出　　处：《肿瘤影像学》2016年第4期-,共5页Oncoradiology

基　　金：国家自然科学基金资助项目(No:81371577;81571676)

摘　　要：目的:制备靶向血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR2)超声造影剂,评价其物理性质及对肿瘤细胞的寻靶能力。方法:利用生物素-亲和素桥接法将微米级超声造影剂USphere与VEGFR2抗体结合,制备靶向VEGFR2超声造影剂,检测其粒径分布;用免疫荧光法检测不同细胞株(高转移肝肿瘤细胞MHCC-97H及正常肝细胞LO2)中VEGFR2表达强度,以及靶向与非靶向VEGFR2超声造影剂对MHCC-97H和LO2细胞的结合能力。结果:靶向VEGFR2超声造影剂微泡分布均匀,平均粒径(1 012.67±78.59)nm,免疫荧光法显示VEGFR2抗体可特异性连接于微泡表面。标记荧光的VEGFR2抗体在MHCC-97H细胞高表达,在LO2细胞低表达。免疫荧光定量显示靶向VEGFR2造影剂在MHCC-97H细胞中的荧光强度比值(微泡荧光强度/细胞荧光强度)为0.75±0.32,显著高于其他3组(P<0.01)。结论:本研究成功制备靶向VEGFR2超声造影剂,其大小稳定均一,可与高表达VEGFR2的MHCC-97H细胞特异性结合。Objective:To evaluate the targeting ability of vascular endothelial growth factor receptor 2 (VEGFR2)-targeted microbubbles to tumor cells and its preparation process.Methods:VEGFR2-targeted microbubbles were accomplished by biotin-avidin linkage of VEGFR2 antibody and USphere ultrasonic contrast agent. The features were detected by dynamic light scattering (DLS) and laser scanning confocal microscope (LSCM). The VEGFR2 expression intensities in MHCC-97H cells (high metastatic human hepatic carcinoma cells) and LO2 cells (human liver cells) were compared by immunolfuorescence method. The combination rates of VEGFR2-targeted microbubbles and non-targeted micro bubbles with both cell lines were documented by quantitative immunofluorescence technique.Results:VEGFR2-targeted microbubbles were distributed evenly with average particle size of (1 012.67±78.59) nm. Immunolfuorescence assay showed green lfuorescence visible on the surface of the targeted micro bubbles. The immunolfuorescence signal was higher in MHCC-97H cells than that in LO2 cells. The lfuorescence signal ratio of VEGFR2-targeted microbubbles was (0.75±0.32) in MHCC-97H cells, which was signiifcantly higher than that in other three groups (P<0.01). Conclusion:VEGFR2-targeted microbubbles with a stable average size are successfully prepared. The micro bubbles could speciifcally combind with VEGFR2-high-expressing MHCC-97H cells.

关 键 词：靶向超声造影剂 生物素-亲和素 血管内皮生长因子受体2 超声分子成像 肿瘤血管生成 

分 类 号：R445.1[医药卫生—影像医学与核医学]