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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 targeted cationic liposomes (CLPs) delivering miRNAs on specific tumor suppressor genes associated with triple negative breast cancer (TNBC). Methods The physical and chemical properties were evaluated by thin-film dispersion method to optimize the cationic lipid composition. A novel hyaluronic acid (HA) CLP with ideal characterization was prepared to characterize the particle size and zeta potential of CLPs. CLPs were surface-modified with HA and the ability of HA-CLPs to enter cells was evaluated by confocal laser scanning and fluorescence detection by flow cytometry. The biological effects of HA-CLPs loaded with miRNAs on cells were studied by EdU cell proliferation assay and cell scratch assay. Results The dispersions of CLPs prepared by the method of membrane dispersion were good, with uniform morphology and hollow spheres. The mean diameter of CLPs was (180.6 ± 3.4) nm and the average zeta potential was (43.4 ± 2.8) mV. The mean diameters of the nanoparticles after HA modification and the nanoparticles carrying miR-205 were (236.65 ± 6.9) nm and (205.6 ± 2.2) nm, respectively, and the mean potentials were (29.1 ± 5.4) mV and ) mV. After confocal laser scanning and flow cytometry fluorescence detection, it was demonstrated that HA-CLPs could successfully deliver miR-205 into the cell and distributed near the nucleus, and the targeting effect of HA was significant. EdU experiments and cell scratch assays demonstrated that miR-205 and miR-34a, which are loaded into TNBC cells, inhibit their proliferation and migration. CONCLUSION: Targeted cationic liposomes have strong targeting specificity to TNBC cells. Targeted delivery of miR-205 and miR-34a can inhibit the proliferation and migration of TNBC cells.