国际生物医学工程杂志
國際生物醫學工程雜誌
국제생물의학공정잡지
INTERNATIONAL JOURNAL OF BIOMEDICAL ENGINEERING
2012年
2期
100-102,后插3
,共4页
蓝瑞隆%张海玲%刘兰霞%冷希岗
藍瑞隆%張海玲%劉蘭霞%冷希崗
람서륭%장해령%류란하%랭희강
双肽修饰壳聚糖%纳米粒%内吞途径
雙肽脩飾殼聚糖%納米粒%內吞途徑
쌍태수식각취당%납미립%내탄도경
Two peptides modified chitosan%Nanoparticle%Endocytic pathway
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摄入的抑制作用高于对TLCDN摄入的抑制作用,但其差异无统计学意义；Filipin明显抑制HepG2对TLCDN的摄入,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.
目的 探索對肝癌細胞HepG2具有較高轉染效率和靶嚮性的TAT-LHRH脩飾的殼聚糖/DNA納米粒的內吞途徑.方法 採用複凝集法,將熒光標記的質粒DNA與殼聚糖或TAT-LHRH脩飾的殼聚糖混閤製備殼聚糖/DNA納米粒(CDN)和TAT-LHRH脩飾的殼聚糖/DNA納米粒(TLCDN).觀察3種內吞途徑抑製劑Chlorpromazine、Filipin或Dynasore對HepG2細胞攝入CDN或TLCDN的影響,用高內涵採集數據併分析.結果 Chlorpromazine對CDN攝入的抑製作用高于對TLCDN攝入的抑製作用,但其差異無統計學意義；Filipin明顯抑製HepG2對TLCDN的攝入,但對CDN的攝入則起促進作用；Dynasore能同時抑製HepG2對上述2種納米粒的攝入,且程度基本相同.結論 肝癌細胞HepG2對CDN的攝入主要通過網格蛋白依賴性細胞內吞途徑,而對TLCDN的攝入途徑包含網格蛋白依賴性細胞內吞途徑和小窩蛋白介導的細胞內吞途徑,但以後者為主.
목적 탐색대간암세포HepG2구유교고전염효솔화파향성적TAT-LHRH수식적각취당/DNA납미립적내탄도경.방법 채용복응집법,장형광표기적질립DNA여각취당혹TAT-LHRH수식적각취당혼합제비각취당/DNA납미립(CDN)화TAT-LHRH수식적각취당/DNA납미립(TLCDN).관찰3충내탄도경억제제Chlorpromazine、Filipin혹Dynasore대HepG2세포섭입CDN혹TLCDN적영향,용고내함채집수거병분석.결과 Chlorpromazine대CDN섭입적억제작용고우대TLCDN섭입적억제작용,단기차이무통계학의의；Filipin명현억제HepG2대TLCDN적섭입,단대CDN적섭입칙기촉진작용；Dynasore능동시억제HepG2대상술2충납미립적섭입,차정도기본상동.결론 간암세포HepG2대CDN적섭입주요통과망격단백의뢰성세포내탄도경,이대TLCDN적섭입도경포함망격단백의뢰성세포내탄도경화소와단백개도적세포내탄도경,단이후자위주.
Objective To explore the endocytic pathway of TAT-LHRH modified chitosan/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 chitosan to form chitosan/DNA nanoparticle (CDN) and TLCDN by the complex coacervation method.Internalization of TLCDN or CDN by HepG2 cells were measured in the presence of three kinds of inhibitors of endocytic 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 nanoprticles.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.
