TAT-LHRH修饰的壳聚糖/DNA纳米粒的细胞内吞途径

目的 探索对肝癌细胞HepG2具有较高转染效率和靶向性的TAT-LHRH修饰的壳聚糖/DNA纳米粒的内吞途径.方法 采用复凝集法,将荧光标记的质粒DNA与壳聚糖或TAT-LHRH修饰的壳聚糖混合制备壳聚糖/DNA纳米粒(CDN)和TAT-LHRH修饰的壳聚糖/DNA纳米粒(TLCDN).观察3种内吞途径抑制剂Chlor...

Endocytic pathways involved in the uptake of TAT-LHRH modified chitosan/DNA nanoparticles by HepG2 cells

Objective To explore the endocytic pathway of TAT-LHRH modified ehitosan/DNA nanoparticle （TLCDN） that exhibits high transfection efficiency and targeting to HepG2. Methods Plasmid DNA was labeled with fluorescein, and the resulting fluorescent DNA was complexed with chitosan or TAT-LHRH modified ehitosan to form chitosardDNA nanoparticle （CDN） and TLCDN by the complex eoacervation method. Internalization of TLCDN or CDN by HepG2 cells were measured in the presence of three kinds of inhibitors of endoeytic pathway, Chlorpromazine, Filipin or Dynasore, using High-Content Analyzer to collect and analyze the data. Results Chlorpromazine led to more decreased uptake of CDN than that of TLCDN, although not statistically significant. Filipin demonstrated significant inhibitory effect on the uptake of TLCDN while promoted the uptake of CDN. Dynasore resulted in a similar decrease in the uptake of both nanoprtieles. Conclusion It was demonstrated that CDN was taken up by HepG2 cells mainly through the clathrin-dependent endocytic pathway and TLCDN was more likely to be internalized by HepG2 cells through the caveolin-mediated endocytic pathway although the clathrin-dependent endocytic pathway was also involved.