靶向纳米造影剂RGD-USPIO标记裸鼠宫颈癌移植瘤MR成像研究  

Study on MR Imaging of Cervical Cancer Xenografts in Nude Mice Using Targeted Nanocontrast Agent RGD-USPIO

作　　者：刘灿 李媛 孙锐 刘东伯 张依玲 漆辉雄[5] LIU Can;LI Yuan;SUN Rui;LIU Dong-bo;ZHANG Yi-ling;QI Hui-xiong(Department of Oncology,Wuhan Fourth Hospital,Wuhan,Hubei,430033,China;Department of Obstetrics and Gynecology,Wuhan Fourth Hospital,Wuhan,Hubei,430033,China;Cancer Center,Renmin Hospital of Wuhan University,Wuhan,Hubei,430060,China;Cancer Center,Tongji Hospital of Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei,430030,China;Department of Oncology,Xiangyang Central Hospital,Affiliated Hospital of Hubei University of Arts and Science,Xiangyang,Hubei,441000,China)

机构地区：[1]武汉市第四医院肿瘤科,湖北武汉430033 [2]武汉市第四医院妇产科,湖北武汉430033 [3]武汉大学人民医院肿瘤中心,湖北武汉430060 [4]华中科技大学同济医学院附属同济医院肿瘤中心,湖北武汉430030 [5]湖北文理学院附属医院、襄阳市中心医院肿瘤科,湖北襄阳441000

出　　处：《现代生物医学进展》2025年第6期984-991,共8页Progress in Modern Biomedicine

基　　金：2021年度武汉市卫生健康委员会科研项目(WX21C28);武汉市科技局2022年度知识创新专项基础研究项目(2020020801010559);国家自然科学基金项目(82372918)。

摘　　要：目的:制备一种针对肿瘤外周新生血管内皮αvβ_(3)整合素的靶向纳米造影剂:精氨酸-甘氨酸-天冬氨酸(RGD)-超小型超顺磁性氧化铁纳米颗粒(USPIO)(RGD-USPIO),并评估其在裸鼠宫颈癌移植瘤磁共振(MR)成像中的标记能力。方法:通过结合RGD小分子环肽(cRGDfE)与USPIO,制备出RGD-USPIO纳米造影剂;测定造影剂的粒径;利用原子吸收分光光度计检测人脐静脉内皮细胞(HUVECS)对RGD-USPIO摄取量的变化。随后,培养宫颈癌Hela细胞株,并将其接种于裸鼠右大腿腹侧皮下。当肿瘤直径达到约8 mm时,对两组裸鼠(实验组和对照组,每组6只)分别通过尾静脉途径注射USPIO或RGD-USPIO造影剂。在造影剂注射之前、注射后0.5 h和2 h,利用4.7T MR进行T2加权扫描,以监测组织T2信号变化。MR扫描结束后处死裸鼠,并采集肿瘤组织进行切片及免疫组化分析,了解不同部位组织的新生血管表达情况。结果:RGD-USPIO的粒径约为15±5 nm,展现出良好的分散度。与USPIO相比,HUVECS在不同时间段对RGD-USPIO的摄取量较高,且该摄取过程可被游离RGD竞争性抑制。注射RGD-USPIO后,实验组肿瘤组织周边的T2驰豫时间明显缩短,免疫组化结果显示肿瘤周边新生血管丰富。结论:RGD-USPIO展现出良好的靶向性和特异性。肿瘤边缘布满活跃的新生血管,这些血管富含整合素αvβ_(3)。利用RGD-USPIO静脉造影技术与4.7T MR扫描相结合,可以精确针对这些新生血管进行定位,为宫颈癌的诊断与治疗开辟了新的途径。Objective:To prepare a targeted nano-contrast agent,arginine-glycine-aspartic acid(RGD)-ultrasmall superparamagnetic iron oxide nanoparticles(USPIO)(RGD-USPIO),for targetingαvβ_(3) integrins in the neovasculature of tumors,and to evaluate its labeling ability in magnetic resonance(MR)imaging of cervical cancer xenografts in nude mice.Methods:RGD-USPIO nano-contrast agent was prepared by conjugating the RGD small molecular cyclic peptide(cRGDfE)with USPIO.The particle size of the contrast agent was determined.Changes in the uptake of RGD-USPIO by human umbilical vein endothelial cells(HUVECS)were detected using an atomic absorption spectrophotometer.Subsequently,cervical cancer Hela cell lines were cultured and inoculated subcutaneously into the right thigh ventral side of nude mice.When the tumor diameter reached approximately 8 mm,USPIO or RGD-USPIO contrast agents were injected via the tail vein into two groups of nude mice(experimental and control groups,6 mice per group).Prior to contrast agent injection,and at 0.5 h and 2 h post-injection,4.7T MR was utilized to perform T2-weighted scans to monitor changes in tissue T2 signals.After the completion of MR scanning,the nude mice were sacrificed,and tumor tissues were collected for sectioning and immunohistochemical analysis to understand the expression of neovasculature in different parts of the tissues.Results:The particle size of RGD-USPIO was approximately 15±5 nm,exhibiting good dispersibility.Compared with USPIO,the uptake of RGD-USPIO by HUVECS was higher at different time points,and the uptake process can be competitively hindered by the presence of free RGD.Following the administration of RGD-USPIO,the T2 relaxation time around the tumor tissue in the experimental group was significantly shortened,and immunohistochemical results showed abundant neovascularization around the tumor.Conclusion:RGD-USPIO exhibited good targeting and specificity.The edges of tumors are densely populated with active neovascularization,which are rich in integrinαvβ_(3).By

关 键 词：宫颈癌 磁共振分子成像 超小型超顺磁性氧化铁粒子 RGD 整合素αvβ_3 

分 类 号：R3[医药卫生—基础医学] R737.3