PD-L1抗体纳米颗粒的制备及其肺癌靶向治疗作用  

Preparation of PD-L1 Antibody Nanoparticles and Its Targeting Therapy for Lung Cancer

作　　者：吴猛[1] 王晓洁[1] 王磊[1] 吕晓文 WU Meng;WANG Xiaojie;WANG Lei;LV Xiaowen(School of Life Sciences,Ludong University,Yantai 264039,China)

机构地区：[1]鲁东大学生命科学学院,山东烟台264039

出　　处：《鲁东大学学报（自然科学版）》2018年第2期144-149,191,192,共7页Journal of Ludong University:Natural Science Edition

基　　金：山东省重点研发计划(2016GNC111016)

摘　　要：制备程序性死亡配体^(-1)(programmed death ligang^(-1),PD-L1)抗体纳米颗粒并对它的表征、载药能力和靶向作用进行了评价,本文利用双乳法和薄膜分散法合成载盐酸阿霉素脂质—聚合物纳米颗粒(简称纳米颗粒),并将其与PD-L1抗体偶联(简称PD-L1抗体纳米颗粒).体外检测纳米颗粒对A549细胞的细胞毒性,流式细胞仪和共聚焦实验检测细胞对纳米颗粒的摄取能力;建立裸鼠A549肺癌肿瘤模型,通过活体成像对PD-L1抗体纳米颗粒的靶向作用和治疗效果进行了评价.结果显示:合成的纳米颗粒粒径集中在100~200 nm,对盐酸阿霉素包封率为60.24%,载药量为5.62%;MTS实验显示非载药纳米颗粒组细胞存活率与正常组无差异,10μg·m L^(-1)和20μg·m L^(-1)浓度的阿霉素纳米颗粒组细胞存活率显著低于同浓度的游离阿霉素组(P<0.05);流式实验显示10μg·m L^(-1)和20μg·m L^(-1)阿霉素浓度的PD-L1抗体纳米颗粒组在瘤细胞中荧光亮度明显高于同浓度的纳米颗粒组;动物实验显示PD-L1抗体纳米颗粒组瘤体积显著小于PBS(磷酸盐缓冲液)对照组和游离阿霉素组(P<0 05).结果表明该合成的PD-L1抗体纳米颗粒,粒度均匀集中、载药能力强,对肺癌具有一定的靶向治疗作用.PD-L1(programmed death ligang-1)antibody nanoparticles were prepared and evaluated for their characterization,drug-loading capacity and targeting.In this paper,double-milk method and film dispersion method were used to synthesize doxorubicin hydrochloride-loading-Lipid-polymer nanoparticles(abbreviated as nanoparticles)and coupled with PD-L1 antibody(abbreviated as PD-L1 antibody nanoparticles).In vitro detection of cytotoxicity of nanoparticles on A549 cells,flow cytometry and the Confocal experiment were used to test the ability of cells to take up nanoparticles.The lung cancer model of mouse A549 was established in vivo,and the targeting and therapeutic effects of PD-L1 antibody nanoparticles were evaluated by in vivo imaging.The results showed that synthetic nanoparticle size concentration in 100~200 nm,the concentration of doxorubicin hydrochloride was 60.24%and the drug loading was 5.62%.The MTS experiments showed that there were no difference in cell survival rate between the non-loaded nanoparticles group and the normal group,the viability of the doxorubicin nanoparticle group at the10μg·mL-1 and 20μg·mL-1 concentration was significantly lower than that of the free doxorubicin group at the same concentration(P<0.05);The flow cytometry showed PD-L1 antibody nanoparticle group at the concentration of 10μg·mL-1 and 20μg·mL-1,the fluorescence intensity of the tumor cells was significantly higher than that of the nanoparticle group with the same concentration;the tumor volume of the PD-L1 antibody nanoparticle group was significantly smaller than that of the PBS(phosphate buffer solution)control group and the free doxorubicin group(P<0.05).The results showed that the synthesized PD-L1 antibody nanoparticles have uniform particle size,strong drug-loading ability,and a certain targeted therapeutic effect on lung cancer.

关 键 词：PD-L1抗体 纳米颗粒 肺癌 靶向治疗 

分 类 号：Q939.91[生物学—微生物学]