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叶酸靶向anti-miR-221阴离子脂质体的制备及体外抗肿瘤作用 Δ

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张文典 ，崔 杰 ，夏一帆 ，段少峰 [1.广东省人民医院（广东省医学科学院）药学部，广州 510080；2.河南大
学药学院，河南 开封 475001]
中图分类号 R943 文献标志码 A 文章编号 1001-0408（2022）07-0813-05
DOI 10.6039/j.issn.1001-0408.2022.07.08

摘 要 目的 基于阴离子脂质体制备一种叶酸靶向miR-221反义寡核苷酸（anti-miR-221）递送系统，并初步评价其体外抗肝癌效
果。方法 采用薄膜分散水化法制备叶酸靶向anti-miR-221脂质体（FRL），测定其粒径、Zeta电位和包封率。以钙黄绿素为模型药
物，通过体外细胞摄取实验观察所制叶酸靶向阴离子脂质体在人肝癌HepG2细胞中的靶向递送效果。利用流式细胞术检测FRL
对 HepG2 细胞凋亡和周期的影响。结果 所制 FRL 的粒径为（172.70±3.76）nm，Zeta 电位为（－1.16±0.15）mV，包封率为
（83.53±1.85）％。体外细胞摄取实验结果显示，叶酸靶向阴离子脂质体成功将模型药物钙黄绿素递送至HepG2细胞中，且递送
效率高于普通非靶向脂质体（P＜0.01）。细胞凋亡检测结果显示，FRL作用后细胞的凋亡率显著高于普通非靶向脂质体作用后的
细胞（P＜0.01）。细胞周期检测结果显示，FRL可使细胞的S期缩短，并将细胞阻滞于G0/G1、G2/M期。结论 FRL可以较好地包封
anti-miR- 221，并成功将其递送至肝癌HepG2细胞中，而且在诱导细胞凋亡和细胞周期调控方面呈现出良好的体外抗肝癌效果。
关键词 miR-221反义寡核苷酸；叶酸；阴离子脂质体；肝癌；靶向；纳米给药系统

Preparation of folate-targeted anti-miR-221 anionic liposome and its in vitro anti-hepatocellular carcinoma
effect
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ZHANG Wendian ，CUI Jie ，XIA Yifan ，DUAN Shaofeng [1. Dept. of Pharmacy，Guangdong Provincial People’s
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Hospital （Guangdong Academy of Medical Sciences），Guangzhou 510080，China；2. School of Pharmacy，
Henan University，Henan Kaifeng 475001，China]
ABSTRACT OBJECTIVE To prepare folate-targeted miR-221 antisense oligonucleotide（anti-miR-221）delivery system，and to
preliminarily evaluate its in vitro anti-cancer effect on hepatocellular carcinoma. METHODS Folate-targeted anti-miR-221
liposomes（FRL）were prepared by thin-film dispersion method；the particle size，Zeta potential and encapsulation efficiency were
determined. The delivery efficiency of folate-targeted anionic liposome in human hepatoma HepG2 cells was determined by in vitro
cellular uptake experiment using calcein as the model drug. Flow cytometry was used to detect the effects of FRL on the apoptosis
and cell cycle of HepG2 cells. RESULTS The particle size of prepared FRL was（172.70±3.76）nm，Zeta potential was（－1.16±
0.15） mV and encapsulation efficiency was （83.53 ± 1.85）％ . In vitro cellular uptake experiments showed that folate-targeted
anionic liposome successfully delivered calcein to HepG2 cells，and the delivery efficiency in targeted group was higher than that of
non-targeted liposome group（P＜0.01）. Apoptosis experiment results showed that the apoptotic rate of HepG2 cells treated with
FRL was significantly higher than that of non-targeted liposome（P＜0.01）. In cell cycle experiment，FRL could shorten the S
phase fraction of HepG2 cells and induced arrest in the G0/G1 and G2/M phases. CONCLUSIONS FRL can encapsulate
anti-miR-221 well and deliver it to hepatocellular carcinoma HepG2 cells successfully，and has a good in vitro anti-hepatoma effect
in inducing apoptosis and cell cycle regulation.
KEYWORDS miR-221 antisense oligonucleotide；folate；anionic liposome；hepatocellular carcinoma；targeted；nanoparticle
delivery system

[1]
在中国，肝癌新发和死亡人数均占全球该疾病患者 期 。目前，被批准用于晚期肝癌的一线治疗药物仅有
的 50％以上，给中国社会和医疗体系带来了沉重的负 索拉非尼和仑伐替尼2种，且其不良反应和耐药现象常
担；且肝癌隐匿性强，超 70％肝癌患者确诊时已处于晚 有报道，通常患者用药数月后会出现肝癌的复发和进
[2]
Δ 基金项目：河南省医学科技攻关计划项目（No.2018020306）；开 展，治疗后生存率和预后较差 。因此，探索新的治疗药
封市科技发展计划项目（No.1903024） 物和治疗策略仍为肝癌治疗的迫切问题。
＊主管药师，硕士。研究方向：靶向纳米给药系统。E-mail：
基因调控理论的发展为癌症等严重疾病的治疗提
zhangwendian@gdph.org.cn
供了新思路。microRNA（后文简称“miRNA”）是一类由
# 通信作者：教授，博士。研究方向：新材料与新型给药系统。
E-mail：sduan@henu.edu.cn 20～24 个核苷酸组成的内源性单链小 RNA 分子，可通

中国药房 2022年第33卷第7期 China Pharmacy 2022 Vol. 33 No. 7 ·813 ·