期刊文献+

构建HMGB1/PEI非病毒载体介导高效基因传递的研究 被引量:2

High Mobility Group B1-Polyethylenimine as Combined Carrier for Gene Delivery in Vitro
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摘要 研究了以聚乙烯亚胺(PEI)为骨架复合高迁移率族蛋白B1(HMGB1)的复合型载体HMGB1/PEI的性能,以期提高非病毒基因载体的转染效率。透射电镜观察pDNA/HMGB1/PEI复合物粒子形态呈球形;动态光散射法测定粒径与表面电位,结果显示复合HMGB1后,复合物粒径降低,且随HMGB1加入量的增大表面电位有增大的趋势;凝胶电泳阻滞试验表明HMGB1可协助PEI与pDNA结合;MTT试验结果显示HMGB1/PEI复合载体的细胞毒性低于PEI;HMGB1/PEI复合载体的转染率较PEI的转染率增大2.9~4.0倍,且HMGB1可以弱化血清对转染的阻碍作用。所以HMGB1被证实能有效提高PEI的体外转染效率。 This study investigates the properties of a combined carrier, which is comprised of polyethylenimine (PEI) and a nuclear peptide, high mobility group box 1 (HMGB1), in order to obtain higher transfection efficiency of non-viral carriers. The pDNA/HMGB1/PEI combined polyplexes are spherical particles observed by transmission electron microscopy. Compared with using PEI alone, improved HMGB1 complexation is observed for smaller particle size of polyplexes and higher surface charge by dynamic light scattering. Gel electrophoresis retardation assays show that HMGB1 could help PEI to bind and condense pDNA. The results of cell viability by MTT assays suggest lower cytotoxicity for HMGB1/ PEI than PEI. The transfection efficiencies for pDNA/HMGB1/PEI combined polyplexes show more than 2. 9 -4. 0 times higher than for pDNA/PEI polyplexes, moreover, HMGB1 could weaken the inhibitory effects of serum on complexes. Thus, the combined carrier HMGB1/PEI which consists of the cationic polymer and the nuclear peptide is proved to enhance transfection efficiency of PEI in vitro.
出处 《中山大学学报(自然科学版)》 CAS CSCD 北大核心 2009年第5期91-96,共6页 Acta Scientiarum Naturalium Universitatis Sunyatseni
基金 国家自然科学基金资助项目(30600789) 广东省自然科学基金资助项目(5300666)
关键词 聚乙烯亚胺 HMGB1 基因传递 polyethylenimine HMGB1 gene delivery
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参考文献14

  • 1NEU M, FISCHER D, KISSEL T. Recent advances in rational gene transfer vector design based on poly (ethylene imine) and its derivatives [J]. J Gene Med, 2005, 7(8) : 992 - 1009.
  • 2LU Y, LOW P S. Folate-mediated delivery of macromolecular anticancer therapeutic agents [ J ]. Adv Drug Deliv Rev, 2002, 54 (5) : 675 - 693.
  • 3KLEEMANN E, NEU M, KELEL N, et al. Nano-carriers for DNA delivery to the lung based upon a TAT-derived peptide covalently coupled to PEG-PEI [ J ]. J Control Release, 2005, 109(1 -3): 299-316.
  • 4HABERLAND A, BOTTGER M. Nuclear proteins as genetransfer vectors [ J]. Biotechnol Appl Biochem, 2005, 42(Pt 2) : 97 - 106.
  • 5BOUSSIF O, LEZOUALCH F, ZANTA M A, et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine [ J ]. Proc Natl Acad Sci, 1995, 92(16) : 7297 -7301.
  • 6GHARWAN H, WIGHTMAN L, KIRCHEIS R, et al. Nonviral gene transfer into fetal mouse livers (a comparison between the cationic polymer PEI and naked DNA) [J]. Gene Ther, 2003, 10(9) : 810 -817.
  • 7TANG M X, SZOKA F C. The influence of polymer structure on the interactions of cationic polymers with DNA and morphology of the resulting complexes [ J ]. Gene Ther, 1997, 4(8) : 823 -832.
  • 8CHOOSAKOONKRIANG S, LOBO B A, KOE G S, et al. Biophysical characterization of PEI/DNA complexes [J]. JPharmSci, 2003, 92(8): 1710-1722.
  • 9ANDERSSON U, ERIANSSON-HARRIS H, YANG H, et al. HMGB1 as a DNA-binding cytokine [J]. J Leuk Biol, 2002, 72: 1084- 1091.
  • 10DUNLAP D D, MAGGI A, SORIA M R, et al. Nanoscopic structure of DNA condensed for gene delivery [J]. Nucleic Acids Res, 1997, 25(15) : 3095 -3101.

二级参考文献22

  • 1Schaffer DV, Fidelman NA, Dan N, et al. Vector unpacking as a potential barrier for receptor-mediated polyplex gene delivery [J].Biotechnol Bioeng, 2000,67(5) : 598 - 606 .
  • 2Wu GY, Wu CH. Receptor-mediated in vitro gene transformation by a soluble DNA carrier system [J]. J Biol Chem, 1987, 262(10):4429 - 4432.
  • 3Brown MD, Schatzlein A, Brownlie A, et al. Preliminary characterization of novel amino acid based polymeric vesicles as gene and drug delivery agents [J]. Bioconjug Chem, 2000, 11(6):880- 891.
  • 4Zinselmeyer BH, Mackay SP, Schatzlein AG, et al. The lowergeneration polypropylenimine dendrimers are effective gene-transfer agents [J]. Pharm Res, 2002, 9(7) :960 - 967.
  • 5Chen Q R, Zhang L, Stass SA, et al. Branched co-polymers of histidine and lysine are efficient carriers of plasmids [J]. Nucleic Acids Res, 2001, 29(6) :1334 - 1340.
  • 6Boussif O, Lezoualc'h F, Zanta MA, et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo:polyethylenimine [J]. Proc Natl Acad Sci USA, 1995, 92(16):7297 - 7301.
  • 7Putnam D, Gentry CA, Pack DW, et al. Polymer-based gene delivery with low cytotoxicity by a unique balance of side-chain termini [J]. Proc Natl Acad Sci U S A,2001, 98(3): 1200 -1205.
  • 8Mosmann T. Rapid colofimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays [ J ]. J Inmmnol Methods, 1983, 65( 1 - 2) :55 - 63.
  • 9Kircheis R, Wightman L, Wagner E. Design and gene delivery activity of modified polyethylenimines [J]. Adv Drug Deliv Rev,2001, 53:341 - 358.
  • 10Godbey WT, Barry, MA, Saggau P, et al. Poly(ethylenimine)mediated transfection: a new paradigm for gene delivery [J]. J Biomed Mater Res, 2000, 51(3) :321 - 328.

共引文献15

同被引文献62

  • 1孟艳,刘嘉茵,王淑玉.细胞周期蛋白及相关蛋白因子在卵母细胞成熟过程中的调控网络[J].国外医学(生理病理科学与临床分册),2004,24(3):267-270. 被引量:1
  • 2钟建辉,银巍,藏林泉,江伟健,邱鹏新,颜光美.腺病毒载体介导的大鼠DNA多聚酶β重组体的构建及其在PC12细胞中的表达[J].中山大学学报(医学科学版),2005,26(6):622-625. 被引量:2
  • 3赵春玲,宋咏梅,樊飞跃,詹启敏.细胞周期蛋白cyclin B1与肿瘤[J].肿瘤,2007,27(4):322-326. 被引量:17
  • 4BAKES J,GORDA S.Genetic improvement of commoncarp strains using intraspecific hybridization[J].Aqua-culture,1995,129:183-186.
  • 5LINHART O,GELA D,RODINA M,et al.Top-crossingwith paternal inheritance testing of common carp(Cypri-nus carpio L.)progeny under two altitude conditions[J].Aquaculture,2002,204:481–491.
  • 6RINDERNECHT E,HUMBEL R E.The amino acid se-quence of human insulin like growth factor 1 and its struc-tural homology with proinsulin[J].J Biol Chem,1978,253(8):2769-2776.
  • 7VASILATOS Y R,SCANES CG.Growth hormone and in-sulin-like growth factor in poultry growth:required,opti-mal or ineffective[J].Poultry Sci,1991,70:1764-1780.
  • 8FLORINI J R,EWTON D Z,MCWADE F J.IGFs,muscle growth,and myogenesis[J].Diabetes Rev,1995,3:73-92.
  • 9SHAMBLOTT M J,CHEN T T.Identification of a secondinsulin-like growth factor in a fish species[J].Proc NatlAcad Sci USA,1992,89:8913–8917.
  • 10DUGUAY S J,LAI-ZHANG J,STEINER D F,et al.Developmental and tissue-regulated expression of IGF-Iand IGF-II mRNAs in Sparus aurata[J].J Mol Endocri-nol,1996,16:123–132.

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