近红外嵌膜共轭寡聚电解质的设计、合成及其在乳腺癌靶向成像中的应用

Design and synthesis of near-infrared embedded conjugated oligoelectrolytes and their application in breast cancer-targeted imagining

目的:探究近红外嵌膜共轭寡聚电解质(COE)在乳腺癌靶向成像中的应用。方法:合成COE-BBT-2T;紫外分光光度计、荧光分光光度计表征COE-BBT-2T的光学性能;CCK-8实验测试COE-BBT-2T的细胞毒性;制备Lip-NK-COE;激光粒度分析仪测试Lip-NK-COE的粒径与电位;透射电镜拍摄脂质体形态;Westernblot验证提取的NK细胞膜是否有效嵌入脂质体中;激光共聚焦显微镜验证Lip-NK-COE对乳腺癌细胞(4T1细胞)是否具有体外靶向性;动物实验分组为COE组、Lip-COE组与Lip-NK-COE组,通过近红外活体成像仪检测Lip-NK-COE在乳腺癌小鼠体内的靶向性和肿瘤成像效果。结果:成功合成COE-BBT-2T分子,该分子具有近红外成像性能和良好的生物相容性;成功制备Lip-NK-COE,粒径为(101.7±2.1)nm,多分散指数为0.224±0.040;提取出的NK细胞膜保留DNAM-1、NKG-2D肿瘤靶向相关蛋白,并有效地嵌入脂质体膜上;通过激光共聚焦显微镜检测出Lip-NK-COE对4T1细胞的靶向性强于Lip-COE;通过近红外活体成像仪检测出Lip-NK-COE对肿瘤的体内靶向性强于Lip-COE。结论:Lip-NK-COE能有效靶向乳腺癌并对其进行成像。

Objective:To explore the application of near-infrared embedded conjugated oligoelectrolytes(COE)in the targeted imaging of breast cancer.Methods:Synthesize COE-BBT-2T and characterize the optical properties of COE-BBT-2T using UV spectrophotometer and fluorescence spectrophotometer;Cytotoxicity experiment(CCK8 experiment)was done to test the cytotoxicity of COE-BBT-2T;Prepare Lip-NK-COE;Laser particle size analyzer was used to test the particle size and potential of Lip-NK-COE;Transmission electron microscopy was used to capture the morphology of liposomes;Western blot was used to verify whether the extracted NK cell membrane was effectively embedded in liposomes,and laser confocal microscopy to verify whether Lip-NK-COE liposomes had the ability to target breast cancer cells(4T1 cells)in vitro;Animal experiments were divided into COE group,Lip-COE group and Lip-NK-COE group to detect the targeting and tumor imaging effect of Lip-NK-COE in breast cancer mice by near-infrared imaging in vivo.Results:The successfully synthesized COE-BBT-2T molecule shows near-infrared imaging performance and good biocompatibility.Lip-NK-COE was successfully prepared,the particle size being(101.7±2.1)nm,and the polydispersity index 0.224±0.040.The extracted NK cell membrane retained DNAM-1,NKG-2D cancer targeting related proteins,which was effectively embed into the liposome membrane.The targeting of Lip-NK-COE and 4T1 cells was detected to be stronger than that of Lip-COE through laser confocal microscopy,and the targeting of tumors in the Lip-NK-COE stronger than that in the Lip-COE using a near-infrared live imaging instrument.Conclusion:Lip-NK-COE can effectively target and image breast cancer.