以凝溶胶蛋白为靶点的超声造影剂对小鼠腹水型肝癌高淋巴转移细胞株摄取的影响  

Effect of ultrasound contrast agent targeting gelatin on uptake of mouse ascites hepatocellular carcinoma cell lines with high lymphatic metastasis

作　　者：何垚[1] 杨龙[1] 袁建军[1] 朱好辉[1] 邵黎阳 He Yao;Yang Long;Yuan Jianjun;Zhu Haohui;Shao Liyang(Department of Ultrasound,Henan People′s Hospital,Zhengzhou 450000,China)

机构地区：[1]河南省人民医院超声科,郑州450000

出　　处：《中华肿瘤杂志》2020年第4期319-324,共6页Chinese Journal of Oncology

基　　金：河南省医学科技攻关计划联合共建项目(2018020431)。

摘　　要：目的:探讨以凝溶胶蛋白为靶点的超声造影剂对小鼠腹腔积液型肝细胞癌高淋巴转移细胞株摄取的影响。方法:采用改良的双乳化溶剂挥发法构建高分子造影剂PLGA-Cooh、碳二亚胺法连接凝溶胶蛋白单抗和造影剂PLGA-Cooh,建立靶向超声造影剂Gsn-PLGA。采用激光粒径仪检测靶向超声造影剂的粒径和Zeta电位,采用免疫荧光法分析造影剂与凝溶胶蛋白单抗表面结合的情况。培养小鼠腹腔积液型肝细胞癌高淋巴转移细胞株Hca-F细胞,采用体外摄取实验分析体外寻靶能力,采用体外显影实验分析造影剂体外显影效果。结果:PLGA-Cooh造影剂呈白色粉末状,具有良好的水溶性,粒径为(569.68±6.96)nm,表面电位为(-10.95±2.43)mV。经DiI标记的非靶向超声造影剂荧光抗体结合率为0.84%,经DiI标记的靶向超声造影剂荧光抗体结合率为95.89%。体外显影实验显示,靶向超声造影剂Gsn-PLGA体外显影效果良好。体外摄取实验显示,与低表达凝溶胶蛋白的Hca-P细胞比较,细胞表面高表达凝溶胶蛋白的Hca-F细胞对靶向超声造影剂的摄取能力较强( P<0.05),可呈现更强的绿色荧光,靶向超声造影剂Gsn-PLGA对凝溶胶蛋白靶点具有较强的靶向性。靶向超声造影剂Gsn-PLGA体外显影实验显示,造影剂回声均匀细腻,后方回声未出现衰减,同时随着造影剂浓度的升高,其显影效果更加明显( P<0.05)。 结论:以凝溶胶蛋白为靶点的超声造影剂Gsn-PLGA可结合高表达凝溶胶蛋白的Hca-F细胞,同时具有良好的体外显影效果。Objective To analyze the effect of ultrasound contrast agent targeting gelatin on uptake of high lymphatic metastasis cell lines of mouse hepatocellular carcinoma with peritoneal effusion.Methods The modified double emulsifying solvent evaporation method was used to construct the macromolecule contrast agent PLGA-Cooh.The carbodiimide was used to connect the monoclonal antibody of gelatin with the contrast agent PLGA-Cooh,and the targeted ultrasound contrast agent Gsn-PLGA was established.The particle size and Zeta potential of the targeted ultrasound contrast agent were measured by laser particle size analyzer.The surface binding of the contrast agent to the gelatin monoclonal antibody was evaluated by immunofluorescence.Hca-F cells with high lymphatic metastasis were cultured in mice with peritoneal effusion hepatocellular carcinoma.Target-seeking ability in vitro was evaluated by in vitro uptake test,and the imaging effect of the contrast agent in vitro was evaluated by in vitro developing test.Results The contrast agent is white powder with good water solubility.The average particle size and surface potential were(569.68±6.96)nm and(-10.95±2.43)mV,respectively.The fluorescent antibody binding rate of non-targeted and targeted ultrasound contrast agent labeled with DiI were 0.84%and 95.89%,respectively.The results showed that the targeted ultrasound contrast agent Gsn-PLGA had a better of developing effect in vitro.Hca-F cells with high expression of gelsolin protein had stronger uptake ability of targeted ultrasound contrast agent and stronger green fluorescence in vitro than those with low expression of gelsolin protein(P<0.05).Moreover,targeted ultrasound contrast agent Gsn-PLGA had stronger targeting to the gelsolin protein.The echo of the targeted ultrasound contrast agent Gsn-PLGA was uniform and fine,without attenuating echo of the back.Simultaneously,the development effect was more obvious with the increase of contrast agent concentration(P<0.05).Conclusion Ultrasound contrast agent Gsn-PLGA targetin

关 键 词：肝肿瘤 超声造影剂 凝溶胶蛋白 聚乳酸羟基乙酸 淋巴转移 

分 类 号：R735[医药卫生—肿瘤]