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TDL复合物提高人脐静脉内皮细胞基因转染效率的体外实验 被引量：1

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摘要 目的研究HIV-1Tat蛋白转导域/质粒DNA/Liposome(TDL)复合物介导增强型绿色荧光蛋白(enhanced green fluorescent protein,pEGFP)在体外培养人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)的转染效率。方法将质粒DNA与Tat肽以不同的电荷比混匀,再加入2μl Lipofectamine2000制成TDL复合物。琼脂糖凝胶电泳分析质粒DNA与Tat肽的结合力;荧光显微镜和流式细胞仪观察TDL复合物与lipofectamine2000介导基因在体外培养人脐静脉内皮细胞中的转染效果;MTT法检测TDL复合物对人脐静脉内皮细胞生长活性的影响。结果TDL复合物组琼脂糖凝胶电泳未发现明显DNA条带。TDL复合物的转染率优于单用Lipofectamine2000(P<0.05)。Tat/DNA电荷比为8∶1时,TDL复合物的转染效率最高。TDL复合物组对细胞活性均无明显影响。结论TDL复合物可明显提高基因在体外培养人脐静脉内皮细胞的转染效率,该方法可作为提高基因转染效率的有效手段之一。
作者 任建丽 王志刚 李兴升 苏琳 张勇 汪朝霞 杨春江 张群霞 许川山
机构地区 重庆医科大学附属第二医院超声科超声影像学研究所
出处 《第三军医大学学报》 CAS CSCD 北大核心 2007年第21期2097-2099,共3页 Journal of Third Military Medical University
基金 国家自然科学基金重点项目(30430230) 国家高技术研究发展规划"863"项目(2006AA02Z4F0)~~
关键词 TDL复合物 人脐静脉内皮细胞 基因转染
分类号 R394-33 [医药卫生—医学遗传学]
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参考文献14

  • 1Madry H, Reszka R, Bohlender J, et al. Efficacy of cationic liposomemediated gene transfer to mesangial cells in vitro and in vivo[ J]. J Mol Med, 2001, 79(4): 184-189.
  • 2Rudolph C, Plank C, Lausier J, et al. Oligomers of the arginine-rich motif of the HIV-1 TAT protein are capable of transferring plasmid DNA into cells[J]. JBiolChem, 2003,278(13): 11411-11418.
  • 3Ignatovich I A, Dizhe E B, Pavlotskaya A V, et al. Complexes of plasmid DNA with basic domain 47 - 57 of the HIV-1 Tat protein are transferred to mammalian cells by endocytosis-mediated pathways[J]. J Biol Chem, 2003,278 (43) : 42625 -42636.
  • 4Vires E, Brodin P, Lebleu B. A truncated HIV-I Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus[ J ]. J Biol Chem, 1997, 272(25 ) : 16010- 16017.
  • 5Liu Z, Li M, Cui D, et al. Macro-branched cell-penetrating peptide design for gene delivery [ J ]. J Control Release, 2005, 102 (3) : 699 -710.
  • 6Miyazaki M, Obata Y, Abe K,et al. Gene transfer using nonviral delivery systems [J]. Perit Dial Int, 2006, 26(6) : 633 -640.
  • 7Alton E W, Stem M, Farley R, et al. Cationic lipid-mediated CFTR gene transfer to the lungs and nose of patients with cystic fibrosis: a double-blind placebo-controlled trial[J]. Lancet, 1999, 353(9157): 947 - 954.
  • 8Tung C H, Mueller S, Weissleder R. Novel branching membrane translocational peptide as gene delivery vector[ J]. Bioorg Med Chem, 2002, 10(11) : 3609 -3614.
  • 9Nakanishi M, Eguchi A, Akuta T, et al. Basic peptides as functional components of non-viral gene transfer vehicles [ J ]. Curr Protein Pept Sci, 2003, 4(2) : 141 - 150.
  • 10Hellgren I, Gorman J, Sylven C. Factors controlling the efficiency of Tat-mediated plasmid DNA transfer[J]. J Drug Target, 2004, 12 (1):39-47.

同被引文献16

  • 1Rebane A,Akdis C A. MicroRNAs:Essential players in the regulation of inflammation[J].Journal of Allergy and Clinical Immunology,2013,(01):15-26.
  • 2van Empel V P,De Windt L J,da Costa Martins P A. Circulating miRNAs:reflecting or affecting cardiovascular disease[J].Current Hypertension Reports,2012,(06):498-509.
  • 3Kim V N,Mithrophanous K,Kingsman S M. Minimal requirement for a lentivirus vector based on human immunodeficiency virus type 1[J].Journal of Virology,1998,(01):811-816.
  • 4Kafri T,van Praag H,Gage F H. Lentiviral vectors:regul-ated gene expression[J].Molecular Therapy,2000,(06):516-521.
  • 5Williams A E,Perry M M,Maschos S A. Role of miRNA-146a in the regulation of the innate immune response and cancer[J].Biochemical Society Transactions,2008,(Pt 6):1211-1215.
  • 6Fontana L,Sorrentino A,Condorelli G. Role of microRN-As in haemopoiesis,heart hypertrophy and cancer[J].Biochemical Society Transactions,2008,(Pt 6):1206-1210.
  • 7Nury D,Neri T,Pueéat M. Human embryonic stem cells and cardiac cell fate[J].Journal of Cellular Physiology,2009,(03):455-459.
  • 8Van Rooij E. The art of microRNA research[J].CIRCULATION RESEARCH,2011.219-234.
  • 9Takaya T,Nishi H,Horie T. Roles of microRNAs and myocardial cell differentiation[J].Prog Mol Biol Transl Sci,2012,(01):139-152.
  • 10Finnegan E J,Matzke M A. The small RNA wodd[J].Journal of Cell Science,2003,(Pt23):4689-4693.

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第三军医大学学报
2007年 第21期

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