C60-IONP-GE11纳米粒对乳腺癌细胞的体外成像及光动力学研究  

Study on C60-IONP-GE11 nanoparticles for MRI imaging and photodynamic therapy on breast cancer cells in vitro

作　　者：符舒琦 涂蓉[1] 刘梦婵 洪雅敏 李韩建[1] 尤晓光 FU Shu-qi;TU Rong;LIU Meng-chan(Department of Radiology,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院放射科,海口570102 [2]海南医学院第二附属医院,海口570216 [3]江西省中医院,南昌330006

出　　处：《放射学实践》2022年第6期715-722,共8页Radiologic Practice

基　　金：2019年海南省科技厅省重点研发计划项目(ZDYF2019200);国家自然科学基金项目(81760310);海南医学院科学培育基金项目(HYYFYPY202014)。

摘　　要：目的:构建多功能MRI对比剂C60-IONP-GE11,研究其体外细胞磁共振靶向成像效果及光动力学治疗效果。方法:免疫荧光染色验证EGFR高表达肿瘤细胞株;体外磁共振成像观察靶向组对比剂C60-IONP-GE11和非靶向组对比剂C60-IONP-RP的成像效果,普鲁氏蓝染色观察对比剂分布情况;CCK8法检测C60-IONP-GE11的细胞毒性;光动力学实验设置单纯激光组、C60-IONP-GE11无激光组、非靶向对比剂+激光组和实验组靶向对比剂C60-IONP-GE11+激光组4个处理组,相应处理后CCK8测其细胞存活率,活性氧检测实验观察不同组ROS产生水平,ImageJ半定量分析ROS荧光水平。结果:免疫荧光实验结果证实MD-MBA-231细胞膜上大量表达EGFR;C60-IONP-GE11体外MRI靶向实验可见T_(2)WI呈负性强化,且T_(2)WI信号随着靶向对比剂浓度增加而降低,线性回归方程为Y=145.898-30.269*X,R2=0.862;普鲁士蓝染色发现C60-IONP-GE11实验组可使MD-MBA-231细胞膜区明显蓝染,而非靶向对照组不能;体外毒性实验发现C60-IONP-GE11无明显细胞毒性;C60-IONP-GE11体外光动力学杀伤乳腺癌细胞,细胞存活率仅为(22.79±4.84)%,并可见细胞内大量活性氧生成,活性氧水平随C60-IONP-GE11浓度增加而增多。结论:C60-IONP-GE11多功能MR对比剂具有体外靶向乳腺癌细胞MR成像功能,几乎无毒,外加激光可靶向杀伤肿瘤细胞,同时具有MRI成像及PDT治疗作用。Objective:To construct a multifunctional MR contrast agent C60-IONP-GE11 and investigate its MR targeted imaging effect and photodynamic therapy effect in vitro.Methods:Immunofluorescence staining was used to confirm the EGFR overexpression tumor cell lines.In vitro MR imaging was used to observe the imaging effect of the targeted group with contrast agent C60-IONP-GE11 and the non-targeted group with contrast agent C60-IONP-RP.And the distribution of the contrast agent was observed by Prussian blue staining.The cytotoxicity of C60-IONP-GE11 was detected by CCK8.Photodynamic experiments were set up in four groups:the laser alone group,C60-IONP-GE11 non-laser group,non-targeted contrast agent+laser group and targeted contrast agent C60-IONP-GE11+laser group.CCK8 assay was used to evaluate cell survival rate after corresponding treatment,and reactive oxygen species(ROS)production was observed in different groups,which can be semi quantitative analysis by Image J.Results:Immunofluorescence confirmed that EGFR was highly expressed on MD-MBA-231 cell membrane.The in vitro MRI targeting experiment of C60-IONP-GE11 showed that T_(2)WI was negatively enhanced,and the T_(2)WI signal decreased with the increase of the concentration of targeting contrast agent.The linear regression equation was Y=145.898-30.269*X,R2=0.862.Prussian blue staining found that C60-IONP-GE11 experimental group could make MD-MBA-231 cell membrane area blue staining,while targeted control group could not.In vitro toxicity tests showed that C60-IONP-GE11 had no obvious cytotoxicity.C60-IONP-GE11 photokinetically killed breast cancer cells in vitro,and the cell survival rate was only(22.79±4.84)%.A large number of reactive oxygen species were generated in the cells,and reactive oxygen species increased with the increase of C60-IONP-GE11 concentration.Conclusion:C60-IONP-GE11 is a multifunctional contrast agent that can target breast cancer cell for MR imaging in vitro,which is almost non-toxic.External laser can target and kill tumor cells.C60-IONP-

关 键 词：富勒烯 氧化铁磁性纳米颗粒 磁共振成像 光化学疗法 

分 类 号：R916.3[医药卫生—药学] R318.08