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凋亡素融合蛋白在HeLa细胞中的转导及体外活性分析 被引量：1

Apoptin Fusion Protein Transduced in HeLa Cells and Its Activity Assay in vitro
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摘要 研究胞外超氧化物歧化酶(EC-SOD)的肝素结合域(HBD)在HeLa细胞中的转导活性,并证实HBD具有运载药物蛋白(凋亡素)进入细胞并诱导肿瘤细胞凋亡的能力。通过克隆表达的HBD-EGFP融合蛋白经Ni2+-NTA纯化后,观察其转导活性;并利用HBD-凋亡素融合蛋白表达纯化后,用噻唑蓝(MTT法)检测肿瘤细胞的凋亡。结果表明:用荧光显微镜可观察到HeLa细胞内有绿色荧光;MTT法检测表明凋亡素融合蛋白能诱导HeLa细胞的凋亡;HBD具有转导活性,能携带药物蛋白(凋亡素)进入细胞并能诱导HeLa细胞的凋亡。 To study transduction activity of the heparin-binding domain（HBD）of extracellular superoxide dismutase（EC-SOD）in HeLa cell,and to confirm HBD have the ability to deliver pharmaceutical proteins（apoptin） into cells and to induce apoptosis of tumor cells.Cloned HBD-EGFP fusion protein was expressed and purified by Ni^（2＋）-NTA affinity chromatography,to observe its transduction activity.HBDapoptin fusion protein was expressed and purified,then the apoptosis of tumor cells in vitro was tested by MTT assay.The results show that green fluorescence can be observed in HeLa cells using fluorescence microscopy.MTT results indicated that fusion protein of apoptin can induce apoptosis of HeLa cells.HBD with the transduction activity,was capable of carrying drugs protein（apoptin） to enter cells and can induce apoptosis in HeLa cells.
作者 华迪 赵健 范立强 王富军 汪雅雯
机构地区 华东理工大学生物反应器工程国家重点实验室 浙江日升昌药业有限公司
出处 《华东理工大学学报（自然科学版）》 CAS CSCD 北大核心 2010年第5期651-654,共4页 Journal of East China University of Science and Technology
关键词 穿膜肽 EC-SOD HELA细胞 cell-penetrating peptides EC-SOD HeLa cell
分类号 Q789 [生物学—分子生物学]
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参考文献17

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同被引文献13

  • 1陈菁,傅蓉,刘树滔,何火聪,饶平凡.Tat蛋白转导区域位于融合蛋白C端时的跨膜递送作用[J].中国生物化学与分子生物学报,2005,21(4):459-464. 被引量：7
  • 2Bitler B G, Schroeder J A. Anti-cancer therapies that utilize cell penetrating peptides[J]. Recent Patents on Anti cancer Drug Discovery,2010,5(2) :99 -108.
  • 3Bolhassani A. Potential efficacy of cell-penetrating peptides for nucleic acid and drug delivery in cancer[J]. Biochimica Et Biophysica Aeta-Reviews on Cancer, 2011,1816(2) : 232-246.
  • 4Koren E, Torchilin V P. Cell penetrating peptides: Breaking through to the other side[J]. Trends in Molecular Medicine, 2012,18(7) :385-393.
  • 5Sawant R, Torehilin V. Intraeellular transduction using cell penetrating peptides[J]. Molecular Biosystems, 2010,6 ( 4 ) : 628-640.
  • 6Grdisa M. The delivery of biologically active (therapeutic) peptides and proteins into cells[J]. Current Medicinal Chem- istry,2011,18(9) :1373-1379.
  • 7Brooks H, Lebleu B, Vives E. Tat peptide-mediated cellular delivery: Back to basics [ J]. Advanced Drug Delivery Reviews,2005,57(4) : 559- 577.
  • 8Adachi Tetsuo, Kodera Tsutomu, Ohta Hideki. The heparin binding site of human extracellular superoxide dismutase[J]. Archives of Biochemistry and Biophysics,1992,297(1) : 155- 161.
  • 9Petersen Steen V, Oury Tim D, Valnickova Zuzana, etaf. The dual nature of human extraeellular superoxide dismutase: One sequence and two structures[J]. Proceedings of the National Academy of Sciences,2003,100(24) :13875-13880.
  • 10Zhang Guohong, Gurtu Vanessa, Kain Steven R. An enhanced green fluorescent protein allows sensitive detection of gene transfer in mammalian cells [J ]. Biochemical and Biophysical Research Communications, 1995, 227 ( 3 ) : 707- 711.

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华东理工大学学报（自然科学版）
2010年 第5期

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