携RGD靶向肽载紫杉醇高分子超声造影剂制备及其对乳腺癌细胞靶向能力、抗肿瘤效应评估  

Preparation of a polymer ultrasound contrast agent loaded with paclitaxel carrying RGD peptide and evaluation of its targeting ability and anti-tumor effect on breast cancer cells

作　　者：李娟[1] 刘桂荣[1] 秦颢诚 武俊 LI Juan;LIU Guirong;QIN Haocheng;WU Jun(The Second People's Hospital of Lianyungang,Lianyungang 222000,China;不详)

机构地区：[1]连云港市第二人民医院,江苏连云港222000 [2]连云港市第一人民医院 [3]大连医科大学附属第二医院

出　　处：《山东医药》2021年第17期5-8,共4页Shandong Medical Journal

基　　金：国家自然科学基金项目(81501476)。

摘　　要：目的制备携精氨酸—甘氨酸—天门冬氨酸(RGD)肽段的RGDfK载紫杉醇(PTX)高分子超声造影剂,并评价其体外乳腺癌细胞靶向能力及抗肿瘤效应。方法用普通造影剂聚乳酸—羟基乙酸(PLGA)为成膜材料,采用双乳化法及碳二亚胺法制备靶向载紫杉醇造影剂PTX-PLGA-RGDfK。光镜下观察该造影剂的形态,马尔文激光粒径仪检测其粒径分布及表面电位,高效液相色谱法测定PTX体外释药能力。联合使用激光共聚焦显微镜及流式细胞仪检测该造影剂表面FITC-RGDfK的连接情况,激光共聚焦显微镜下观察体外对乳腺癌细胞靶向情况,CCK-8法检测乳腺癌细胞存活率评价细胞毒性,超声诊断仪造影模式下观察其显影效果。结果造影剂PTX-PLGA-RGDfK水溶性较好、大小较均一,粒径为(326.24±56.63)nm,表面电位为-11.4 mV;PTX包封率为82.10%±2.12%,载药量为8.21%±0.21%。PTX-PLGA-RGDfK在最初的12 h内药物突释,释放率达46.59%±1.83%;随后出现平稳的缓慢释放,72 h后释放率达73.26%±1.98%。该造影剂表面FITC-RGDfK结合率达95.28%±0.37%,乳腺癌细胞周围可见大量红色荧光靶向造影剂。与PLGA同浓度处理的乳腺癌细胞比较,PTX-PLGA、PTX-PLGA-RGDfK处理的乳腺癌细胞存活率降低,且PTX-PLGA-RGDfK的乳腺癌细胞处理低于PTX-PLGA处理者(P均<0.05)。PTXPLGA-RGDfK体外显影成像效果较好,回声均匀细腻。结论成功制备出携RGD肽载PTX的高分子超声造影剂,可与MBA-MD-231细胞特异性结合并提高化疗药物对肿瘤细胞的杀伤效果。Objective To prepare the RGDfK-carrying arginine-glycine-aspartate RGD peptide(RGD peptide)polymer ultrasound contrast agent containing paclitaxel(PTX),and to evaluate its ability to target breast cancer cells and its anti-tumor effect in vitro.Methods The common contrast agent polylactic acid-glycolic acid(PLGA)was used as the film-forming material,and the targeted drug-loaded paclitaxel contrast agent PTX-PLGA-RGDfK was prepared by the double emulsification method and the carbodiimide method.The morphology of the contrast agent was observed under a light microscope,the particle size distribution and surface potential were detected by a Malvern laser particle sizer,and the PTX in vitro release ability was determined by high performance liquid chromatography.The confocal laser microscope and flow cytometer were used to detect the connection of FITC-RGDfK on the surface of the contrast agent.The targeting effect on breast cancer cells in vitro was observed under the laser confocal microscope,and the survival rate of breast cancer cells was detected by CCK-8 to evaluate its cytotoxicity.We observed the imaging effect of the ultrasound diagnostic apparatus in the contrast mode.Results The contrast agent PTX-PLGA-RGDfK had good water solubility in a uniform size,with a particle size of(326.24±56.63)nm and a surface potential of-11.4 mV;the PTX encapsulation efficiency was 82.1%±2.12%,and the drug loading was 8.21±%0.21%.PTX-PLGA-RGDfK was released in the first 12 h,and the release amount reached 46.59%±1.83%;then there was a steady and slow release,and the release rate reached 73.26%±1.98%after 72 h.The average FITC-RGDfK binding rate on the surface of the contrast agent was 95.28±0.37%,and a large number of red fluorescent targeted contrast agents were seen around breast cancer cells.Compared with the breast cancer cells treated with the same concentration of PLGA,the survival rate of breast cancer cells treated with PTX-PLGA and PTX-PLGA-RGDfK decreased,and the survival rate of breast cancer cells treated wit

关 键 词：RGD肽 αvβ3整合素受体 紫杉醇 超声造影剂 靶向治疗 乳腺癌 

分 类 号：R73-36[医药卫生—肿瘤]