PLGA纳米载体介导的端粒酶反义核酸体外转染率及对肝肿瘤细胞的影响

In vitro transfection efficiency of telomerase antisensenucleic acids carried by PLGA nanoparticles and its influence on tumor cell of liver

目的通过以PLGA纳米粒作为端粒酶hTERT反义核酸的载体,促进反义核酸对肝肿瘤细胞的抑制作用。方法用绿色荧光蛋白报道基因pEGFP-N1表达质粒DNA转染细胞;观测不同转染方法的转染率,TRAP-ELISA法检测端粒酶活性;噻唑蓝(MTT)法测定端粒酶反义核酸对肝肿瘤细胞HepG2的抑制;过氧化物酶的抗荧光素抗体法检测肿瘤细胞凋亡指数。结果裸DNA和磷酸钙的转染效率均不高,分别为18%和24%;脂质体的转染效率为42%,纳米粒的转染效率最高,达61%;以脂质体或PLGA纳米粒为载体介导端粒酶反义核酸处理的肿瘤细胞,其端粒酶活性和生长与空白对照相比明显下降(P<0.05),两种载体之间的差异也非常显著(P<0.05);处理72h后,纳米粒介导组的肿瘤细胞凋亡率显著高于脂质体介导组,分别为23.1%和16.4%。结论PLGA纳米粒是一种非常有前途的非病毒基因治疗载体。

[Objective] To improve inhibition of tumor cell of liver through telomerase antisensenucleic acids carried by PLGA nanoparticles. [Methods] To determine transfection efficiency of different methods by transfection of reporter gene of green fluorescent protein into cells. To measure telomerase activation through TRAP-ELISA. To test influence of telomerase antisensenucleic acids on inhibition of tumor cell of liver by MTr methods. To determine apoptotic index of tumor cell by means of anti-dioxyfluoran antibody of peroxydase. [Result] Transfection efficiency mediated through PLGA nanoparticles was highest, 61%, liposome is next, 42%. Telomerase activation and growth rate of tumor cells treated with antisensenucleic acids decreased, and significant difference existed between two cartiers （P 〈0.05）. 72 h after treatment, apoptotic index of tumor cell in nanoparticle carry group was 23.1%, notably, higher than that in liposome carry group, 16.4%. [Conclusion] PLGA nanoparticle is potential as non-viral carry for gene therapy.