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中华肝脏外科手术学电子杂志 ›› 2018, Vol. 07 ›› Issue (05) : 414 -419. doi: 10.3877/cma.j.issn.2095-3232.2018.05.016

所属专题: 文献

基础研究

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
杨光华1, 张建勋1, 殷保兵2,()   
  1. 1. 200137 上海中医药大学附属第七人民医院普通外科
    2. 200040 上海,复旦大学附属华山医院普通外科;200040 上海,复旦大学附属华山医院静安分院普通外科
  • 收稿日期:2018-06-30 出版日期:2018-10-10
  • 通信作者: 殷保兵
  • 基金资助:
    上海市第七人民医院人才培养计划(QMX2016-01)

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

Guanghua Yang1, Jianxun Zhang1, Baobing Yin2,()   

  1. 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; Department of General Surgery, Jing'an Branch of Huashan Hospital Affiliated to Fudan University, Shanghai 200040, China
  • Received:2018-06-30 Published:2018-10-10
  • Corresponding author: Baobing Yin
  • About author:
    Corresponding author: Yin Baobing, Email:
引用本文:

杨光华, 张建勋, 殷保兵. 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[J]. 中华肝脏外科手术学电子杂志, 2018, 07(05): 414-419.

Guanghua Yang, Jianxun Zhang, Baobing Yin. Inhibitory effect of long-circulating miR-135a-containing cationic immunoliposomes modified by anti-EGFR on gallbladder carcinoma cells[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2018, 07(05): 414-419.

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 anti-EGFR-CIL-pWPXL (LSD-t=37.62, P<0.05). 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.

图1 电镜下经磷钨酸染色后anti-EGFR-CILs粒子形态
图2 荧光显微镜下观察LCL-FITC和anti-EGFR-CIL-FITC转染胆囊癌细胞情况
图3 胆囊癌细胞侵袭能力Transwell实验
图4 胆囊癌细胞凋亡实验
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