精氨酸-甘氨酸-天冬氨酸环肽修饰的纳米粒在小鼠乳腺癌细胞中的靶向性及超声成像研究  

Targeting and CEUS Ability of cRGD Modified Nanoparticles in Breast Cancer Cells

作　　者：申洪远 陶宏宇 张芙蓉 王晨雨 米成嵘[2] SHEN Hongyuan;TAO Hongyu;ZHANG Furong;WANG Chenyu;MI Chengrong(Department of Ultrasound,Ningxia Medical University,Yinchuan 750004,China;不详)

机构地区：[1]宁夏医科大学临床医学院,宁夏银川750004 [2]宁夏医科大学总医院超声科,宁夏银川750004

出　　处：《中国医学影像学杂志》2022年第5期423-429,共7页Chinese Journal of Medical Imaging

基　　金：国家自然科学基金项目(81960317)。

摘　　要：目的制备精氨酸-甘氨酸-天冬氨酸环肽(c RGD肽)修饰的聚乙二醇-聚乳酸-羟基乙酸(PEG-PLGA)高分子聚合物靶向纳米造影剂c RGD-PEG-PLGA@PFPNPs(c RGD-NPs),探究其表征、相变及对小鼠乳腺癌细胞(4T1)的靶向性及超声成像。材料与方法利用双乳化法制成c RGD-NPs,验证c RGD与PEG-PLGA连接情况,观察cRGD-NPs表征、热致相变稳定性等基本特征;检测其对4T1细胞活性的影响;使用激光共聚焦显微镜,将BALB/c小鼠分为靶向组3只及非靶向组3只,通过活体成像观察小鼠体内外乳腺癌细胞摄取c RGD-NPs情况;观察c RGD-NPs声致相变及造影效果。结果成功制备c RGD-NPs,光镜与透射电镜下均表现为形态规则、大小均一的球形,平均粒径为(93.19±31.41)nm;不同浓度c RGD-NPs同4T1细胞共孵育不同时间后,细胞活力均>85%。激光共聚焦显微镜及活体组织荧光成像观察显示,与非靶向纳米粒组相比,c RGD-NPs组更多聚集于肿瘤细胞周围。随着低强度聚焦超声辐照时间推移增加,体外超声造影成像效果越好,在相同条件下也实现了体内乳腺肿瘤显影。结论cRGD-NPs是一种可靶向聚集于4T1细胞区域,具有靶向定位释药潜能且可显影的纳米级载体,可为临床乳腺癌化疗及靶向治疗提供新思路。Purpose To prepare PEG-PLGA targeted nanoparticle contrast agent c RGD-PEG-PLGA@PFP NPs(c RGD-NPs),and observe its characterization,active targeting ability for mouse breast cancer cells(4T1 cells)and imaging effect.Materials and Methods c RGD-NPs were prepared by the double emulsification method.The connection of cRGD and PEG-PLGA,characterization and morphology of NPs were detected and observed.The safety of nanoparticles was detected.Confocal laser microscopy and in vivo imaging in mouse(3 in targeted group,3 in non-targeted group)were used to observe the uptake of targeted nanoparticles in breast cancer cells.The phase transformation and contrast imaging effect of nanoparticles were observed.Results c RGD-NPs was successfully prepared.Under optical microscope and transmission electron microscope,cRGD-NPs showed regular spherical shape and uniform size,with an average particle size of(93.19±31.41)nm.The cell viability of 4T1 cells were all>85%after co-incubation with different concentrations of nanoparticles for different time.Confocal laser microscopy and fluorescence imaging in vivo showed that cRGD-NPs clustered around tumor cells more than those in the non-targeted labeling group.With the increase of low-intensity focused ultrasound irradiation time,the imaging effect of contrast-enhanced ultrasound in vitro was enhanced,and the CEUS imaging of breast tumors in vivo was also achieved under the same conditions.Conclusion c RGD-PEG-PLGA@PFP NPs is a nanoscale carrier that can not only target the 4T1 breast cell region actively,but also realize CEUS imaging,and has potential of targeted drug release,which provides a new idea for clinical breast cancer chemotherapy and targeted therapy.

关 键 词：cRGD肽 纳米造影剂 靶向性 乳腺肿瘤 超声检查 

分 类 号：R445.1[医药卫生—影像医学与核医学] R737.9[医药卫生—诊断学]