【摘要】 目的研究靶向生存素的载小干扰RNA(siRNA)纳米粒对小鼠乳腺癌细胞的体外杀伤作用。方法复凝聚法制备靶向生存素的载siRNA纳米粒,采用动态光散射粒度仪和透射电镜对纳米粒的粒径、电位和形态进行研究;应用划痕实验和CCK-8方法考察载siRNA纳米粒抑制乳腺癌细胞增殖的效果;倒置显微镜和流式细胞仪检测载siRNA纳米粒对4T1细胞凋亡的影响。结果所制备的载siRNA球形纳米粒平均粒径为202.2 nm,多分散系数0.121,Zeta电位为+18.58 mV;划痕实验结果显示,转染48 h后,裸siRNA组由于细胞增殖,划痕愈合,而纳米粒组细胞增殖受到抑制,划痕宽度没有变化;CCK-8实验结果表明,转染后24 h纳米粒对4T1细胞增殖的抑制率可达45.4%,较裸siRNA组(约5.5%)显著增高(P<0.05);凋亡检测结果显示,纳米粒组凋亡细胞达(98.77±0.61)%,与裸siRNA组(6.66±0.44)%相比,有显著统计学差异(P<0.05)。结论以生存素为靶点的载siRNA纳米粒对小鼠乳腺癌细胞有较强的杀伤作用。更多还原

【Abstract】 Objective To evaluate the killing effect of siRNA loaded nanoparticles targeting survivin on mice mammary tumor cells in vitro. Methods Complex coacervation method was used to prepare siRNA loaded nanoparticles. Particle size,Zeta potential and the morphology of nanoparticles were determined by dynamic light scattering and the transmission electron microscopy,respectively; anti-proliferation effects of siRNA loaded nanoparticles was assessed by CCK-8 and the wound healing assay. On the other hand,apoptotic 4T1 cells induced by nanoparticles were observed using an inverted microscope,and the percentage of apoptotic cells were quantified by flow cytometry. Results spherical nanoparticles with particle size of 202. 2 nm and polydispersity index of 0. 121 were prepared,and positive charge of + 18. 58 mV existed on the surface of nanoparticles; results from the wound healing assay showed that the wound disappeared in the naked siRNA group finally,but there were no difference in the size of wound in the chitosan-g-transcriptional activator protein( CTS-g-TAT) siRNA nanoparticles group with the culture time increasing; results from CCK-8 assay showed that inhibition efficiency of CTS-g-TAT / siRNA nanoparticles could reach 45. 4%,which was much higher than that of 5. 5% in the naked siRNA group( P < 0. 05); moreover,the percentage of apoptotic cells nanoparticles was( 98. 77 ± 0. 61) % in the CTS-g-TAT / siRNA group,which was much higher than that of( 6. 66 ± 0. 44) % in the naked siRNA group from the results of flow cytometry( P < 0. 05).Conclusion The siRNA loaded nanoparticles which target survivin had a strong killing effect against mouse mammary tumor cells.更多还原