摘要
目的:探讨羟基磷灰石-聚乙烯亚胺(nHA-PEI 10KD)纳米颗粒的癌细胞基因转染效率。方法:通过透射电子显微镜(TEM)观察HA-PE(I10KD)纳米颗粒的形态及粒径,Zeta电位仪测定nHA-PEI和HA在酸、碱、中性环境中的电位,用琼脂糖凝胶电泳检测nHAP-PE(I10KD)与DNA结合的能力,MTT比色法检测nHAP-PE(I10KD)对HepG2细胞的毒性,选用增强型绿色荧光蛋白质粒pEGFP1与nHA-PEI结合后,分别转染真核细胞HepG2、Hela、SW620,计算其转染率。结果:nHA-PE(I10KD)分散程度好,粒径60-80nm,在PH7.2时,Zeta电位42.87mV,能转染实验中的细胞,转然效果最好的是HepG2细胞,其次Hela、SW620,转染率高于PEI(10KD)、nHA,但低于脂质体。结论:通过阳离子PEI修饰HA,可有效将增强型绿色荧光蛋白质粒转入HepG2细胞,HA-PE(I10KD)纳米颗粒复合物有望成为基因传递的有效载体。
Objective：To investigate the efficiency of gene transfection mediated by hydroxyapatite-polyethyleneimine（nHA-PEI 10KD） nanoparticles in cancer cells.Methods： The shape and size of HA-PEI nano-particle were observed by Transmission Electron Microscopy（TEM）,The potential of nHA-PEI and HA in acid,alkaliand neutral conditions were measured by Zeta Potential Instrument.DNA binding capacity of nHAP-PEI（10KD） was detected by agarose gel electrophoresis,MTT assy was performed to evaluate the toxicity ofnHAP-PEI （10KD） to HepG2 cells. The combinations of enhanced green fluorescent protein plasmid pE（3FP1 and nHA-PEI were transfected into HepG2, Hela and SW620 cells, and the transfection efficiency was evaluated by Flow Cytomea＇y. Remtlts： nHA-PEI （10KD） are in high dispersion and the size of particle is 60 - 80nm. At PH of 7.2, Zeta Potential of 42.87mV, nHA-PEI transfects into experiment cells, especially in HepG2 cells, followed by Hela, SW620.The transfection efficiency ofnHA-PEI was higher than PEI （10） and nHA, but lower than liposomes. Conclusions： The modified HA by PEI cation can potentiate the transfection of enhanced green fluorescent protein plasmid into HepG2 cells. HA-PEI （10KD） composite nanoparticles are expected to become an effective gene delivery vector.
出处
《现代生物医学进展》
CAS
2009年第19期3669-3672,3695,共5页
Progress in Modern Biomedicine
关键词
羟基磷灰石
聚乙烯亚胺
基因载体
转染效率
hydroxyapatite
polyethylenimine
gene vector
transfection efficiency
