叶酸靶向anti-miR-221阴离子脂质体的制备及体外抗肿瘤作用
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篇名: | 叶酸靶向anti-miR-221阴离子脂质体的制备及体外抗肿瘤作用 |
TITLE: | Preparation of folate-targeted anti-miR- 221 anionic liposome and its in vitro anti-hepatocellular carcinoma effect |
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。 |
ABSTRACT: | OBJECTIVE To prep are 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 HepG 2 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 HepG 2 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 HepG 2 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 HepG 2 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 HepG 2 cells and induced arrest in the G 0/G1 and G 2/M phases. CONCLUSIONS FRL can encapsulate anti-miR-221 well and deliver it to hepatocellular carcinoma HepG 2 cells successfully ,and has a good in vitro anti-hepatoma effect in inducing apoptosis and cell cycle regulation. |
期刊: | 2022年第33卷第07期 |
作者: | 张文典,崔杰,夏一帆,段少峰 |
AUTHORS: | ZHANG Wendian ,CUI Jie,XIA Yifan,DUAN Shaofeng |
关键字: | 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 |
KEYWORDS: | miR-221 antisense oligonucleotide ;folate;anionic liposome ;hepatocellular carcinoma ;targeted;nanoparticle |
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