TAT—LHRH修饰的壳聚糖／DNA纳米粒的细胞内吞途径  

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

作　　者：蓝瑞隆[1] 张海玲[1] 刘兰霞[1] 冷希岗[1] 

机构地区：[1]中国医学科学院北京协和医学院生物医学工程研究所,天津300192

出　　处：《国际生物医学工程杂志》2012年第2期100-102,I0003,共4页International Journal of Biomedical Engineering

基　　金：天津市应用基础及前沿技术重点项目（1IJCZDJC20300,09JCZDJC18700）

摘　　要：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Chlorpromazine、Filipin或Dynasore对HepG2细胞摄入CDN或TLCDN的影响，用高内涵采集数据并分析。结果Chlorpromazine对CDN摄人的抑制作用高于对TLCDN摄人的抑制作用，但其差异无统计学意义；Filipin明显抑制HepG2对TLCDN的摄人，但对CDN的摄入则起促进作用；Dynasore能同时抑制HepG2对上述2种纳米粒的摄入，且程度基本相同。结论肝癌细胞HepG2对CDN的摄入主要通过网格蛋白依赖性细胞内吞途径，而对TLCDN的摄入途径包含网格蛋白依赖性细胞内吞途径和小窝蛋白介导的细胞内吞途径，但以后者为主。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.

关 键 词：双肽修饰壳聚糖 纳米粒 内吞途径 

分 类 号：R318.08[医药卫生—生物医学工程]