The Preparation of uPAR-Targeted MRI Probe and its Targetability to Breast Cancer Cells
Yang Yang1, Meng Jie1#, Wen Tao1, Chen Bo2, Liu Fei3, Gu Ning2, Xu Haiyan1#, Yu Wei4*, Liu Jian1#*
1(Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine PekingUnion Medical College, Beijing 100005, China) 2(Jiangsu Key Laboratory for Biomaterials and Devices, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China) 3 (Research Institute of Chia Tai Tianqing Pharmaceutical Group Limited by Share Ltd, Nanjing 210018, China) 4(Department of Radiology, Beijing An Zhen Hospital, Capital Medical University, Beijing 100029, China)
Abstract：Breast cancer is one of the most common malignancies for women. Early detection of primary and metastatic lesions of breast cancer is a key point to improve the survival rate of patients. Studies have shown that urokinase-type plasminogen activator receptor (uPAR) is highly expressed in breast cancer cells and tumor-associated cells, while lowly expressed in normal tissue cells. In this study, by conjugating amino-nitrilotriacetic acid (NTA) to the surface of superparamagnetic iron oxide nanoparticles (SPIO), we obtained an universal magnetic resonance imaging (MRI) module (SPIO-NTA). Then, amino-terminal fragment (ATF) of urokinase-type plasminogen activator (uPA), the native ligand of uPAR, was introduced to SPIO-NTA via histidine tags (His tags) to obtain an uPAR-targeted MRI agent (SPIO-ATF). The diameter of the γ-Fe2O3 core was less than 10 nm. The hydrodynamic diameter of SPIO-ATF was about 77 nm and Zeta potential was about -13 mV. The results of protein gel electrophoresis and Coomassie brilliant blue staining indicated the successful conjugation of ATF to SPIO. In vitro experiments showed a positive correlation between the amount of SPIO-ATF bound with 4T1 cells and the expression level of uPAR, indicating that SPIO-ATF can specifically bind to 4T1 cells via the interaction between ATF and uPAR. In addition, the cellular uptake of SPIO-ATF was increased with the increase of incubating concentration of SPIO-ATF. Under experimental conditions, neither SPIO nor SPIO-ATF showed obvious cytotoxicity. In conclusion, we developed a novel MRI probe for the target detection of breast cancer, providing a new strategy for the early diagnosis of breast cancer.
杨扬, 孟洁, 温涛, 陈博, 刘飞, 顾宁, 许海燕, 于薇, 刘健. uPAR靶向性MRI探针的制备及其与乳腺癌细胞靶向结合的研究[J]. 中国生物医学工程学报, 2018, 37(4): 481-488.
Yang Yang, Meng Jie, Wen Tao, Chen Bo, Liu Fei, Gu Ning, Xu Haiyan, Yu Wei, Liu Jian. The Preparation of uPAR-Targeted MRI Probe and its Targetability to Breast Cancer Cells. Chinese Journal of Biomedical Engineering, 2018, 37(4): 481-488.
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