期刊文献+

NK4基因的克隆及其在Raji细胞中的表达 被引量:5

Cloning and expression of NK4 gene in Raji cells
下载PDF
导出
摘要 目的:克隆NK4基因,构建其重组真核表达载体,并观察其在Raji细胞中的表达。方法:从人肝组织中提取总RNA,行RT-PCR获得NK4基因cDNA,与载体pVITRO2-mcs构建重组真核表达载体,转染Raji细胞。潮霉素B筛选后,采用实时荧光定量PCR、ELISA、细胞免疫组化和半固体培养等方法检测NK4基因在Raji细胞的表达,筛选稳定表达的细胞株。结果:NK4基因获成功克隆并构建了重组真核表达载体pVITRO2-mcs-NK4;转染NK4基因后的Raji细胞经RT-PCR发现存在特异性DNA片段。NK4基因在Raji细胞可稳定表达NK4mR-NA和蛋白,且可抑制Raji细胞的增殖、迁徙和侵袭。结论:NK4基因成功克隆并构建了重组真核表达载体,转染Raji细胞后表达稳定。 AIM: To clone NK4 gene and to construct recombinant eukaryotic expression vector for observing its expression in transfected Raji cells. METHODS: Total RNA was extracted from human hepatic tissue. NK4 gene cDNA was amplified by RT - PCR, and then cloned into vector pVITRO2 - mcs to construct the recombinant eukaryotic expression vector pVITRO2 - mcs - NK4. Raji cells were transfected by recombinant vector pVITRO2 - mcs - NK4 and screened by homomycin B. The stable strain of NK4 gene expression was screened by real - time fluorescent quantitative PCR, ELISA, immunocytohistochemistry and semisolid culture. RESULTS: The specific DNA fragment was detected by RT- PCR in Raji cells transfected with NK4 gene. The transfected Raji cells expressed NK4 mRNA and protein stably, which inhibited Raji cell proliferation, metastasis and invasion. CONCLUSION: NK4 gene is cloned and recombined to construct recombinant eukaryotic expression vector pVITRO2 - mcs - NK4 successfully. NK4 gene in Raji cells expresses stably.
出处 《中国病理生理杂志》 CAS CSCD 北大核心 2009年第5期970-975,共6页 Chinese Journal of Pathophysiology
基金 浙江省医药卫生科学基金资助项目(No2003B172 No.2007A175) 宁波市医药卫生科学基金资助项目(No.2003079) 宁波市科技计划资助项目(No.2007C10065)
关键词 基因 NK4 克隆 基因表达 RAJI细胞 Genes, NK4 Cloning Gene expression Raji cells
  • 引文网络
  • 相关文献

参考文献15

  • 1Kuba K, Matsumoto K, Date K, et al. HGF/NK4, a four - Kringle antagonist of hepatocyte growth factor, is an angiogenesis inhibitor that suppresses tumor growth and metastasis in mice [ J ]. Cancer Res, 2000, 60 (23) :6737 - 6743. 被引量:1
  • 2Jiang WG, Hiscox SE, Parr C, et al. Antagonistic effect of NK4, a novel hepatocyte growth factor variant, on in vitro angiogenesis of human vascular endothelial cells [ J].Clin Cancer Res, 1999, 5(11) :3695 -3703. 被引量:1
  • 3Kanehira M, Xin H, Hoshino K, et al. Targeted delivery of NK4 to multiple lung tumors by bone marrow - derived mesenchymal stem cells [ J ]. Cancer Gene Ther, 2007, 14( 11 ) :894 -903. 被引量:1
  • 4Hiscox S, Parr C, Nakamura T, et al. Inhibition of HGF/SF induced breast cancer cell motility and invasion by the HGF/SF variant, NK4 [ J ]. Breast Cancer Res Treat, 2000, 59(3) :245 -254. 被引量:1
  • 5Saga Y, Mizukami H, Suzuki M, et al. Expression of HGF/NK4 in ovarian cancer ceils suppresses intraperitoneal dissemination and extends host survival [ J ]. Gene Ther, 2001, 8(19) :1450 - 1455. 被引量:1
  • 6Okasora T, Jo JI, Tabata Y. Augmented anti - tumor therapy through natural targetability of macrophages genetically engineered by NK4 plasmid DNA [J]. Gene Ther, 2008, 15(7) :524 -530. 被引量:1
  • 7Du W, Hattori Y, Yamada T, et al. NK4, an antagonist of hepatoeyte growth factor (HGF), inhibits growth of multiple myeloma ceils: molecular targeting of angiogenic growth factor [J]. Blood, 2007, 109(7) :3042 -3049. 被引量:1
  • 8Zou HY, Li Q, Lee JH, et al. An orally available small - molecule inhibitor of c - Met, PF - 2341066, exhibits cytoreductive antitumor efficacy through antiproliferative and antiangiogenic mechanisms [ J ]. Cancer Res, 2007, 67 (9) :4408 -4417. 被引量:1
  • 9岑东,吕建新,裴仁治,涂植光,余晓林,文阳安.肝细胞生长因子mRNA实时荧光定量PCR检测体系的建立及其在淋巴瘤诊断中的价值[J].中华检验医学杂志,2008,31(4):384-388. 被引量:5
  • 10Son G, Hirano T, Seki E, et al. Blockage of HGF/c - Met system by gene therapy (adenovirus -mediated NK4 gene) suppresses hepatocellular carcinoma in mice [ J ]. J Hepatol, 2006, 45 (5) :688 - 695. 被引量:1

二级参考文献31

  • 1方天翎,闵军,邓小耿,钱世鵾,褚忠华,陈亚进,邵静,魏菁,陈积圣.肝细胞生长因子诱导骨髓间质干细胞分化为心肌细胞[J].中国病理生理杂志,2004,20(7):1167-1170. 被引量:18
  • 2Rovers MM,Schilder AG,Zielhuis GA,Rosenfeld RM,张江平,杨妙丽,张全安.中耳炎[J].国外医学(耳鼻咽喉科学分册),2005,29(3):141-143. 被引量:440
  • 3周敦华,魏菁,黄绍良,方建培,包蓉,吴燕峰,黎阳.人脐血来源MSCs的主要生物学特性的研究[J].中国病理生理杂志,2006,22(8):1606-1609. 被引量:7
  • 4Nakamura K, Nishizawa T, Higiya M, et al. Molecular cloning and expression of human hepatoeyte growth factor. Nature, 1989, 342:440-443. 被引量:1
  • 5Forte G, Minieri M, Cossa P, et al. Hepatocyte growth factor effects on mesenchymal stem cells: proliferation, migration and differentiation. Stem Cells, 2006,24:23-33. 被引量:1
  • 6Tjin EP, Groen RW, Vogelzang I, et al. Functional analysis of HGF/MET-signaling and aberrant HGF-activator expression in diffuse large B cell lymphoma. Blood, 2006,107:760-768. 被引量:1
  • 7Pons E, Uphoff C, Drexler HG. Expression of hepatocyte growth factor and its receptor C-met in human leukemia-lymphoma cell lines. Leuk Bes, 1998,22:797-804. 被引量:1
  • 8Gorzelntiak K, Janke J, Engeli S, et al. Validation of endogenous controls for gene express studies in human adipocytes and preadipocytes. Horm Metab Res, 2001,33:625-627. 被引量:1
  • 9Sambrook J, Russell DW. Molecular Cloning: A Laboratory Manual. 3rd ed. New York: Cold Spring Harbor laboratory press, 2001,8:86-112. 被引量:1
  • 10Harting I, Wiesner RJ. Quantification of transcript-to-template rations as a measure of gene expression using RT-PCR. Biotechniques, 1997,23:450-454. 被引量:1

共引文献8

同被引文献31

  • 1Jian-Zheng Jie,Jian-Wei Wang,Jian-Guo Qu,Wei Wang,Tao Hung.Effects of adenoviral-mediated gene transduction of NK4 on proliferation, movement, and invasion of human colonic LS174T cancer cells in vitro[J].World Journal of Gastroenterology,2006,12(25):3983-3988. 被引量:3
  • 2李慧,张农.HGF抑制肾小球硬化作用机制的研究进展[J].临床与实验病理学杂志,2007,23(2):215-217. 被引量:2
  • 3Kuba K,Matsumoto K,Date K,et al.HGF/NK4,a four-Kringle antagonist of hepatocyte growth factor,is an angiogenesis inhibitor that suppresses tumor growth and metastasis in mice[J].Cancer Res,2000,60(23):6737-43. 被引量:1
  • 4Jiang W G,Hiscox S E,Parr C,et al.Antagonistic effect of NK4,a novel hepatocyte growth factor variant,on in vitro angiogenesis of human vascular endothelial cells[J].Clin Cancer Res,1999,5(11):3695-703. 被引量:1
  • 5Kanehira M,Xin H,Hoshino K,et al.Targeted delivery of NK4 to multiple lung tumors by bone marrow-derived mesenchymal stem cells[J].Cancer Gene Ther,2007,14(11):894-903. 被引量:1
  • 6Hiscox S,Parr C,Nakamura T,et al.Inhibition of HGF/SF induced breast cancer cellmotility and invasion by the HGF/SF variant,NK4[J].Breast Cancer Res Treat,2000,59(3):245-54. 被引量:1
  • 7Saga Y,Mizukami H,Suzuki M,et al.Expression of HGF/NK4 in ovarian cancer cells suppresses intraperitoneal dissemination and extends host survival[J].Gene Ther,2001,8(19):1450-5. 被引量:1
  • 8Okasora T,Jo J I,Tabata Y.Augmented anti-tumor therapy through natural targetability of macrophages genetically engineered by NK4 plasmid DNA[J].Gene Ther,2008,15(7):524-30. 被引量:1
  • 9Forte G,Minieri M,Cossa P,et al.Hepatocyte growth factor effects on mesenchymal stem cells:proliferation,migration,and differentiation[J].Stem Cells,2006,24(1):23-33. 被引量:1
  • 10Kim J G,Sohn S K,Kim D H,et al.Clinical implications of angiogenic factors in patients with acute or chronic leukemia:Hepatocyte growth factor levels have prognostic impact,especially in patients with acute myeloid leukemia[J].Leuk Lymphoma,2005,46(6):885-91. 被引量:1

引证文献5

二级引证文献5

;
使用帮助 返回顶部