白蛋白纳米载体介导的反义核酸对肝癌细胞端粒酶活性的抑制

Inhibition on telomerase activity of HepG2 cells by antisense oligonucleotides of telomerase carried by albumin nanoparticles

目的利用纳米载体增强端粒酶反义寡核苷酸对肝脏肿瘤细胞端粒酶活性的抑制作用。方法实验设6组:空白对照组、纯纳米粒组、游离正义寡核苷酸组、游离反义寡核苷酸组、正义寡核苷酸-纳米粒组和反义寡核苷酸-纳米粒组。用TRAP-ELASA法检测端粒酶反义寡核苷酸对肿瘤细胞端粒酶活性的影响,以MTT法测定端粒酶反义寡核苷酸对肝脏肿瘤细胞(HepG2)生长的抑制作用。结果①纯纳米粒组、游离端粒酶正义核苷酸组和端粒酶正义核苷酸-白蛋白纳米粒处理组的端粒酶活性与空白对照组差异无显著性(P>0.05)。②游离端粒酶反义核苷酸和端粒酶反义核苷酸-白蛋白纳米粒处理的肿瘤细胞的端粒酶活性与空白对照相比明显下降(P<0.05),且两者之间差异存在显著性(P<0.05)。③纯纳米粒组、游离端粒酶正义核苷酸和端粒酶正义核苷酸-白蛋白纳米粒对肿瘤细胞的抑制作用很小。游离端粒酶反义核苷酸的抑制作用明显,端粒酶反义核苷酸-白蛋白纳米粒的抑制作用最强,两者之间差异有显著性(P<0.05)。结论白蛋白纳米粒可作为反义技术治疗的有效载体,增强端粒酶反义核苷酸的抑制作用。

[Objective] To investigate the growth inhibition of antisense oligonucleotides of telomerase on HepG2 cells enhanced by albumin nanoparticles.[Methods] There were six groups in this study： blank group, albumin nanoparticles group, free plus sense oligonucleotides group, free antisense oligonucleotides of telomerase group. plus sense oligonucleotides and albumin nanoparticles group, antisense oligonucleotides of telomerase and albumin nanoparticles group. Influence of antisense oligonucleotides of telomerase on telomerase was studied by TRAP-E- LASA method. The inhibition of antisense oligonucleotides of telomerase on HEPG2 growth was studied by MTT method. [Results] Compared with blank group, there are no obvious differences in the telomerase activity of HepG2 in group albumin nanopartieles group, free plus sense oligonucleotides group and plus sense oligonucleotides and albumin nanoparticles group （P 〉0.05）. Compared with blank group blank group, there are obvious differences in the telomerase activity of HepG2 in free antisense oligonucleotides of telomerase group and antisense oligonucleotides of telomerase and albumin nanoparticles group（P 〉0.05）, and the two groups are parently different from each other（P 〉0.05）. The inhibition on HepG2 in albumin nanoparticles group, free plus sense oligonucleotides group, plus sense oligonueleotides and albumin nanoparticles group is negligible, while it is obvious in free antisense oligonucleotides of telomerase group and antisense oligonueleotides of telomerase and albumin nanoparticles group. And the inhition of group antisense oligonucleotides of telomerase and albumin nanoparticles group is the most obvious （P 〉0.05）. [Conclusion] Albumin nanoparticles can be used as an effective carrier in anfisense technology.