Tang Yue, Liu Yun, Xie Yuwen, Chen Jing, Dou Yushun
Department of Pharmacy, China Pharmaceutical University, Nanjing 211198, China.
Asian J Pharm Sci. 2020 Jan;15(1):121-128. doi: 10.1016/j.ajps.2018.09.009. Epub 2018 Nov 27.
Gene-based therapeutics has emerged as a promising approach for human cancer therapy. Among a variety of non-viral vectors, polymer vectors are particularly attractive due to their safety and multivalent groups on their surface. This study focuses on guanidinylated O-carboxymethyl chitosan (GOCMCS) along with poly-β-amino ester(PBAE) for siRNA delivery. Binding efficiency of PBAE/siRNA/GOCMCS nanoparticles were characterized by gel electrophoresis. The siRNA-loaded nanoparticles were found to be stable in the presence of RNase A, serum and BALF respectively. Fine particle fraction (FPF) which was determined by a two-stage impinger (TSI) was 57.8% ± 2.6%. The particle size and zeta potential of the nanoparticles were 153.8 ± 12.54 nm and + 12.2 ± 4.94 mV. cell transfection studies were carried out with A549 cells. The cellular uptake was significantly increased. When the cells were incubated with siSurvivin-loaded nanoparticles, it could induce 26.83% ± 0.59% apoptosis of A549 cells and the gene silencing level of survivin expression in A549 cells were 30.93% ± 2.27%. The results suggested that PBAE/GOCMCS nanoparticle was a very promising gene delivery carrier.
基于基因的疗法已成为一种很有前景的人类癌症治疗方法。在多种非病毒载体中，聚合物载体因其安全性和表面的多价基团而特别具有吸引力。本研究聚焦于胍基化的O-羧甲基壳聚糖（GOCMCS）以及聚-β-氨基酯（PBAE）用于小干扰RNA（siRNA）递送。通过凝胶电泳对PBAE/siRNA/GOCMCS纳米颗粒的结合效率进行了表征。发现负载siRNA的纳米颗粒分别在核糖核酸酶A、血清和支气管肺泡灌洗液存在的情况下是稳定的。通过两级冲击器（TSI）测定的细颗粒分数（FPF）为57.8%±2.6%。纳米颗粒的粒径和zeta电位分别为153.8±12.54nm和+12.2±4.94mV。用A549细胞进行了细胞转染研究。细胞摄取显著增加。当细胞与负载siSurvivin的纳米颗粒孵育时，它可诱导A549细胞26.83%±0.59%的凋亡，并且A549细胞中生存素表达的基因沉默水平为30.93%±2.27%。结果表明PBAE/GOCMCS纳米颗粒是一种非常有前景的基因递送载体。
