Liu Lanxia, Dong Xia, Zhu Dunwan, Song Liping, Zhang Hailing, Leng Xigang G
Laboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People's Republic of China.
Int J Nanomedicine. 2014 Jun 10;9:2879-89. doi: 10.2147/IJN.S61392. eCollection 2014.
To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC). TLC/DNA nanoparticles (TLCDNPs) were characterized by agarose gel retardation, atomic force microscopy, and dynamic light scattering analysis. In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System. The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70-85 nm and zeta potential of +30 mV and were relatively stable in solution. The in vitro study demonstrated TLC was highly selective for hepatoma cells and essentially nontoxic.
为了开发一种能够将基因特异性递送至肝癌细胞的基于壳聚糖的非病毒基因载体，将由TAT（转录反式激活因子）肽和促黄体生成素释放激素（LHRH）组成的双功能肽与低分子量壳聚糖偶联，得到TAT-LHRH-壳聚糖偶联物（TLC）。通过琼脂糖凝胶阻滞、原子力显微镜和动态光散射分析对TLC/DNA纳米颗粒（TLCDNPs）进行表征。使用GE IN Cell Analyzer 2000高内涵细胞分析系统分析体外靶向特异性和转染效率。结果表明，TLC比未修饰的壳聚糖具有更强的DNA凝聚能力，并且TLCDNPs大致呈圆形，平均直径为70-85nm，ζ电位为+30mV，在溶液中相对稳定。体外研究表明，TLC对肝癌细胞具有高度选择性且基本无毒。
