切换至 "中华医学电子期刊资源库"

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/OL]. 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/OL]. 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
[1]
Hundal R, Shaffer EA. Gallbladder cancer: epidemiology and outcome[J]. Clin Epidemiol, 2014(6):99-109.
[2]
Shen HX, Song HW, Xu XJ, et al. Clinical epidemiological survey of gallbladder carcinoma in northwestern China, 2009-2013: 2379 cases in 17 centers[J]. Chronic Dis Transl Med, 2017, 3(1):60-66.
[3]
Garzon R, Marcucci G, Croce CM. Targeting microRNAs in cancer: rationale, strategies and challenges[J]. Nat Rev Drug Discov, 2010, 9(10):775-789.
[4]
Chandra V, Kim JJ, Mittal B, et al. MicroRNA aberrations: an emerging field for gallbladder cancer management[J]. World J Gastroenterol, 2016, 22(5):1787-1799.
[5]
Zhou H, Guo W, Zhao Y, et al. MicroRNA-135a acts as a putative tumor suppressor by directly targeting very low density lipoprotein receptor in human gallbladder cancer[J]. Cancer Sci, 2014, 105(8):956-965.
[6]
Pais-Costa SR, Farah JF, Artigiani-Neto R, et al. Evaluation of P53, E-cadherin, Cox-2, and EGFR protein imunnoexpression on prognostic of resected gallbladder carcinoma[J]. Arq Bras Cir Dig, 2014, 27(2):126-132.
[7]
Kirpotin DB, Noble CO, Hayes ME, et al. Building and characterizing antibody-targeted lipidic nanotherapeutics[J]. Methods Enzymol, 2012(502):139-166.
[8]
Juang V, Lee HP, Lin AM, et al. Cationic PEGylated liposomes incorporating an antimicrobial peptide tilapia hepcidin 2-3: an adjuvant of epirubicin to overcome multidrug resistance in cervical cancer cells[J]. Int J Nanomedicine, 2016(11):6047-6064.
[9]
Ramamoorth M, Narvekar A. Non viral vectors in gene therapy-an overview[J]. J Clin Diagn Res, 2015, 9(1):GE1-6.
[10]
Sanna V, Pala N, Sechi M. Targeted therapy using nanotechnology: focus on cancer[J]. Int J Nanomedicine, 2014(9):467-483.
[11]
Adiseshaiah PP, Hall JB, McNeil SE. Nanomaterial standards for efficacy and toxicity assessment[J]. Wiley Interdiscip Rev Nanomed Nanobiotechnol, 2010, 2(1):99-112.
[12]
Goodwin T, Huang L. Nonviral vectors: we have come a long way[J]. Adv Genet, 2014(88):1-12.
[13]
Shi J, Votruba AR, Farokhzad OC, et al. Nanotechnology in drug delivery and tissue engineering: from discovery to applications[J]. Nano Lett, 2010, 10(9):3223-3230.
[14]
Yin H, Kanasty RL, Eltoukhy AA, et al. Non-viral vectors for gene-based therapy[J]. Nat Rev Genet, 2014, 15(8):541-555.
[15]
Mortensen JH, Jeppesen M, Pilgaard L, et al. Targeted antiepidermal growth factor receptor (cetuximab) immunoliposomes enhance cellular uptake in vitro and exhibit increased accumulation in an intracranial model of glioblastoma multiforme[J]. J Drug Deliv, 2013:209205.
[16]
Kuai JH, Wang Q, Zhang AJ, et al. Epidermal growth factor receptor-targeted immune magnetic liposomes capture circulating colorectal tumor cells efficiently[J]. World J Gastroenterol, 2018, 24(3):351-359.
[17]
Yamada Y, Hidaka H, Seki N, et al. Tumor-suppressive microRNA-135a inhibits cancer cell proliferation by targeting the c-MYC oncogene in renal cell carcinoma[J]. Cancer Sci, 2013, 104(3):304-312.
[18]
Dang Z, Xu WH, Lu P, et al. MicroRNA-135a inhibits cell proliferation by targeting Bmi1 in pancreatic ductal adenocarcinoma[J]. Int J Biol Sci, 2014, 10(7):733-745.
[19]
Shin JY, Kim YI, Cho SJ, et al. MicroRNA 135a suppresses lymph node metastasis through down-regulation of ROCK1 in early gastric cancer[J]. PLoS One, 2014, 9(1):e85205.
[20]
Tang W, Jiang Y, Mu X, et al. MiR-135a functions as a tumor suppressor in epithelial ovarian cancer and regulates HOXA10 expression[J]. Cell Signal, 2014, 26(7):1420-1426.
[1] 杨桂清, 孟静静. 哺乳期亚临床乳腺炎的研究进展[J/OL]. 中华乳腺病杂志(电子版), 2024, 18(06): 376-379.
[2] 戴睿, 张亮, 陈浏阳, 张永博, 吴丕根, 孙华, 杨盛, 孟博. 肠道菌群与椎间盘退行性变相关性的研究进展[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(06): 546-549.
[3] 张洁, 罗小霞, 余鸿. 系统性免疫炎症指数对急性胰腺炎患者并发器官功能损伤的预测价值[J/OL]. 中华普外科手术学杂志(电子版), 2025, 19(01): 68-71.
[4] 赖全友, 高远, 汪建林, 屈士斌, 魏丹, 彭伟. 三维重建技术结合腹腔镜精准肝切除术对肝癌患者术后CD4+、CD8+及免疫球蛋白水平的影响[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(06): 651-654.
[5] 梁孟杰, 朱欢欢, 王行舟, 江航, 艾世超, 孙锋, 宋鹏, 王萌, 刘颂, 夏雪峰, 杜峻峰, 傅双, 陆晓峰, 沈晓菲, 管文贤. 联合免疫治疗的胃癌转化治疗患者预后及术后并发症分析[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(06): 619-623.
[6] 张志兆, 王睿, 郜苹苹, 王成方, 王成, 齐晓伟. DNMT3B与乳腺癌预后的关系及其生物学机制[J/OL]. 中华普外科手术学杂志(电子版), 2024, 18(06): 624-629.
[7] 司钦亮, 毕世龙, 焦慧骁, 李世照, 陈哲禹, 武玉东. 精索去分化脂肪肉瘤两例并文献复习[J/OL]. 中华腔镜泌尿外科杂志(电子版), 2024, 18(06): 585-590.
[8] 王大伟, 陆雅斐, 皇甫少华, 陈玉婷, 陈澳, 江滨. 间充质干细胞通过调控免疫机制促进创面愈合的研究进展[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(06): 361-366.
[9] 刘文竹, 唐窈, 刘付臣. 诱导多潜能干细胞在神经肌肉疾病研究中的应用进展[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(06): 367-373.
[10] 魏志鸿, 刘建勇, 吴小雅, 杨芳, 吕立志, 江艺, 蔡秋程. 肝移植术后急性移植物抗宿主病的诊治(附四例报告)[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(06): 846-851.
[11] 张龙, 孙善柯, 徐伟, 李文柱, 李俊达, 池涌泉, 何广胜, 成峰, 王学浩, 饶建华. 腹腔镜脾切除治疗血液系统疾病的临床疗效分析[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(06): 870-875.
[12] 中华医学会器官移植学分会. 肝移植术后缺血性胆道病变诊断与治疗中国实践指南[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(06): 739-748.
[13] 陈伟杰, 何小东. 胆囊癌免疫靶向治疗进展[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(06): 763-768.
[14] 董佳, 王坤, 张莉. 预后营养指数结合免疫球蛋白、血糖及甲胎蛋白对HBV 相关慢加急性肝衰竭患者治疗后预后不良的预测价值[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(06): 555-559.
[15] 谭瑞义. 小细胞骨肉瘤诊断及治疗研究现状与进展[J/OL]. 中华临床医师杂志(电子版), 2024, 18(08): 781-784.
阅读次数
全文


摘要


AI


AI小编
你好!我是《中华医学电子期刊资源库》AI小编,有什么可以帮您的吗?