EGFR抗体修饰的长循环阳离子免疫脂质体转载miR-135a对胆囊癌细胞的抑制作用  

Inhibitory effect of long-circulating miR-135a-containing cationic immunoliposomes modified by anti-EGFR on gallbladder carcinoma cells

作　　者：杨光华[1] 张建勋[1] 殷保兵[2,3] Yang Guanghu1, Zhang Jianxun1, Yin Baobinge2,3(1.Department of General Surgery, Seventh People＇s Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China;2.Department of General Surgery, Huashan Hospital Affiliated to Fudan University, Shanghai 200040, China;3.Department of General Surgery, Jing＇an Branch of Huashan HospitaI Affiliated to Fudan University, Shanghai 200040, China)

机构地区：[1]上海中医药大学附属第七人民医院普通外科,200137 [2]复旦大学附属华山医院普通外科,上海200040 [3]复旦大学附属华山医院静安分院普通外科,上海200040

出　　处：《中华肝脏外科手术学电子杂志》2018年第5期414-419,共6页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：上海市第七人民医院人才培养计划(QMX2016-01)

摘　　要：目的构建由表皮生长因子受体(EGFR)抗体修饰的长循环阳离子免疫脂质体(anti-EGFR-CILs),评估其理化性质及靶向性,探讨由其转载miR-135a对胆囊癌细胞的侵袭转移及凋亡等生物学行为的影响。方法采用薄膜分散-超声水化法制备转载miR-135a的免疫脂质体,检测其形态、粒径分布、ζ-电位、包封率、载药率、转染率等。通过Transwell侵袭实验、Annexin V/PI双染色法实验评估miR-135a对胆囊癌细胞的侵袭、转移和凋亡的影响。多组吸光度值(A)比较采用单因素方差分析和LSD-t检验。结果 anti-EGFR-CILs具有良好的理化性质及靶向性,包封率和载药率分别为73.91%和1.43%,体外细胞转染率86.5%。Transwell实验显示,与anti-EGFR-CIL-pWPXL相比,anti-EGFR-CIL-miR-135a对胆囊癌细胞具有明显的抑制作用(LSD-t=37.62,P<0.05)。细胞凋亡实验显示miR-135a促进胆囊癌细胞凋亡,细胞凋亡率约21%。结论 anti-EGFR-CILs是一种靶向性强、高效的基因载体。anti-EGFR-CIL-miR-135a可作为一种潜在的、可选择的胆囊癌治疗手段。Objective To construct the long-circulating cationic immunoliposomes modified by anti-epidermal growth factor receptor（EGFR）（anti-EGFR-CILs）, to evaluate the physicochemical properties and targeting ability, and explore its effect of transloading miR-135a on the invasion, metastasis and apoptosis of gallbladder carcinoma（GBC） cells. Methods The immunoliposomes loaded with miR-135a were prepared by film-dispersion and hydration-sonication methods. The morphology, particle size distribution, ζ-potential, drug entrapment efficiency, drug loading efficiency and transfection rate were detected. The effect of miR-135a on the invasion, metastasis and apoptosis of GBC cells were evaluated by Transwell assay and Annexin V/PI staining.The absorbance values（A）among different groups were compared by single factor analysis of variance and LSD-t test.Results anti-EGFR-CILs possessed excellent physicochemical properties and targeting ability.The drug entrapment efficiency and drug loading efficiency of anti-EGFR-CILs were 73.91%and 1.43%.The in vitro cell transfection rate of 86.5%.Transwell assay demonstrated that anti-EGFR-CIL-miR-135a exerted significantly higher inhibitory effect upon GBC cells compared with antiEGFR-CIL-pWPXL（LSD-t=37.62,P0.05）.Cellular apoptosis assay revealed that miR-135a promoted the apoptosis of GBC cells and the apoptosis rate was approximately 21%.Conclusions anti-EGFR-CILs is a highly targeted and efficient gene vector.anti-EGFR-CIL-miR-135a is a potential and alternative therapy for gallbladder cancer.

关 键 词：胆囊肿瘤 受体 表皮生长因子 免疫 脂质体 miR-135a 基因 

分 类 号：R735.8[医药卫生—肿瘤]