主动靶向型载奥拉帕尼PEI-PLGA纳米粒的制备表征及抗三阴乳腺癌研究

Preparation,Characterization and Anticancer Effect of Active-targeted Olaparib-loaded PEI-PLGA Nanoparticles

目的构建具有肿瘤细胞特异靶向性的纳米给药系统,研究该纳米粒的体外抗三阴乳腺癌作用。方法采用乳化扩散.溶剂挥发法制备包载奥拉帕尼的聚乙烯亚胺=聚乳酸-羟基乙酸共聚物(PEI-PLGA)纳米粒,进一步用透明质酸修饰,制备靶向载药纳米粒。采用激光粒度仪和透射电镜对纳米粒的形态、粒径大小、电位及分布情况进行表征。CCK-8法检测纳米粒对MDA-MB-231细胞的毒性作用,通过共聚焦显微镜观察细胞对目标纳米粒的靶向摄取,生物透射电镜观察细胞内部形态变化分析凋亡情况。结果成功制备奥拉帕尼靶向载药纳米粒(HA-Ola-PPNPs),粒径为(166.2±0.842)nm,大小分布均匀,呈圆形或椭圆形,稳定性良好;奥拉帕尼包载于PEI-PLGA纳米粒中能够起到药物缓释作用;CCK-8实验结果显示纳米粒组均呈现出时间依赖和剂量依赖性细胞毒性,且HA-Ola-PPNPs对癌细胞具有更强的杀伤作用;激光共聚焦结果说明了HA与CD44受体的主动靶向结合可以显著提高MDA-MB-231细胞对目标纳米粒的摄取;生物透射电镜观察显示HA-Ola-PPNPs能够有效诱导MDA-MB-231细胞发生凋亡。结论本研究成功制备了具有CD44受体特异性靶向的载药纳米粒,为三阴乳腺癌的临床治疗提供一种新策略。

OBJECTIVE To synthesize the active-targeted olaparib-loaded polyethyleneimine-polyllactic-co-glycolic acid(PEI-PLGA) nanoparticles(HA-Ola-PPNPs) and investigate the therapy of triple negative breast cancer in vitro.METHODS The olaparib-loaded PEI-PLGA nanoparticles were prepared by the emulsion solvent diffusion evaporation method. And further modified with hyaluronic acid(HA) to prepare HA-Ola-PPNPs. The synthesis of HA-Ola-PPNPs was verified by dynamic light scattering(DLS) and transmission electron microscopy(TEM), the cytotoxicity was measured by CCK-8.Confocal microscopy was used to observe the uptake of NPs. Biological TEM was used to observe the morphological changes of cells and analyze the apoptosis.RESULTS The mean size of HA-Ola-PPNPs was(166.2±0.842)nm and showed negatively charged surface(-12.3±1.63)mV. The encapsulation efficiency of HA-Ola-PPNPs was(80.44±6.218)%. CCK-8 results showed that all the nanoparticles presented time-dependent and dose-dependent cytotoxicity, and HA-Ola-PPNPs exhibited efficient in vitro antitumor activities. The binding of HA to CD44 receptor could significantly improve the uptake of HA-Ola-PPNPs by MDA-MB-231 cells.And HA-Ola-PPNPs could effectively induce apoptosis of MDA-MB-231 cells.CONCLUSION In this study, active-targeted HA-Ola-PPNPs are successfully prepared, which could provide a new strategy for the clinical treatment of triple negative breast cancer(TNBC).