靶向阳离子脂质体递送乳腺癌抑制作用的研究miRNA对三阴性  

Delivery of tumor suppressive miRNA to triple negative breast cancer by targeting cationic liposome

作　　者：徐枫[1] 刘婷[1] 赵亚男[2] 陆森[1] 陈园园[1] 叶园园[1] 刘培峰[2] 韩宝三[1] 

机构地区：[1]上海交通大学医学院附属新华医院普外科普通外科实验室,200092 [2]上海交通大学医学院附属仁济医院中心实验室,200127

出　　处：《中华普通外科学文献（电子版）》2017年第3期148-153,共6页Chinese Archives of General Surgery(Electronic Edition)

基　　金：国家自然科学基金资助项目(81172078);宁波市社会发展重大择优科技攻关项目(2012C5013)

摘　　要：目的探讨靶向阳离子脂质体(CLPs)递送miRNA对三阴性乳腺癌(TNBC)相关特异性抑瘤基因的作用。方法通过薄膜分散法评估理化性质对阳离子脂质成分进行优化,制备具有理想表征性质的新型透明质酸(HA)CLP,表征CLPs的粒径和zeta电位。对CLPs进行表面HA靶向修饰,并通过激光共聚焦观察和流式细胞术的荧光检测评估HA-CLPs进入细胞的能力。利用EdU细胞增殖检测和细胞划痕实验对负载miRNA的HA-CLPs对细胞产生的生物学效应进行研究。结果薄膜分散法制备的CLPs纳米颗粒分散性良好、形态均一,呈中空球形结构,CLPs平均粒径为(180.6±3.4)nm、平均zeta电位为(43.4±2.8)mV。经过HA修饰后的纳米颗粒及运载miR-205后的纳米颗粒平均粒径分别为(236.65±6.9)nm、(205.6±2.2)nm,平均电位分别为(29.1±5.4)mV、(11.2±1.1)mV。经过激光共聚焦检测和流式细胞术荧光检测,验证了HA-CLPs能够顺利将miR-205运送至细胞内而分布在细胞核附近,且HA的靶向作用显著。EdU实验和细胞划痕实验验证了运载入TNBC细胞内的miR-205和miR-34a对其增殖和迁移的抑制作用。结论制备的靶向阳离子脂质体对TNBC细胞具有较强的靶向特异性,且通过靶向递送miR-205和miR-34a对TNBC细胞的增殖和迁移起到了一定的抑制作用。Objective To investigate the effect of delivering specific tumor suppressor genes miRNA in triple negative breast cancer （TNBC）.Methods Film dispersion method was utilized to prepare for the new-pattern hyaluronic acid （HA）-targeted cationic liposome （HA-CLPs）. The physicochemical property, such as the representation of the size and zeta-potential of CLPs nanoparticles, was assessed and then the nanoparticles were modified with HA. Confocal and flow cytometry （FCM） was applied to assess targeting capacity, while EdU cell proliferation test and cell wound scratch assay to assess biological effects of HA-CLPs.Results Nanoparticles of CLPs were of good dispersibility and shaped as homogeneous form of hollow globe. The average size of CLPs was （180.6±3.4） nm and the average zeta-potential was （43.4±2.8） mV. The average size of HA-targeted nanoparticles and miRNA-loaded HA-targeted nanoparticles was （236.65±6.9） nm and（205.6±2.2） nm. The average zeta-potential of HA-targeted nanoparticles and miRNA-loaded HA-targeted nanoparticles was （29.1±5.4） mV and （11.2±1.1） mV. Confocal and FCM verified the targeting capacity of HA-CLPs to deliver miR-205 around the MDA-MB-231 cell nucleus observably.EdU cell proliferation test and cell wound scratch assay proved that miR-205 and miR-34a sent into MDA-MB-231 cell had the ability to inhibit cell proliferation and migration.Conclusion Targeted cationic liposome has specific targeting ability towards MDA-MB-231 cell, and the delivery of miR-205 and miR-34a plays a part in the proliferation and migration inhibition of MDA-MB-231.

关 键 词：脂质体 阳离子 透明质酸 乳腺肿瘤 

分 类 号：R737.9[医药卫生—肿瘤]