期刊文献+

转染NK4基因对人胰腺癌SW1990细胞生物学特性的影响 被引量:3

Effect of Transferred NK4 Gene on Biological Characteristics of Human Pancreatic Cancer Cell Line SW1990
下载PDF
导出
摘要 背景与目的:肝细胞生长因子(hepatocytegrowthfactor,HGF)通过其受体c-Met的激活,在调节肿瘤侵袭和转移和血管形成中具有重要作用。NK4不仅是HGF的拮抗剂,也是血管形成的抑制剂,阻断HGF/c-Met途径和肿瘤血管的形成可成为抗肿瘤治疗的策略之一。为此,我们构建NK4基因真核细胞表达载体并进行转染研究,探讨NK4基因在胰腺癌细胞中的表达及其对胰腺癌细胞生物学特性的影响。方法:对重组pcDNA3/hNK4质粒进行酶切,将NK4基因克隆到真核表达载体pRC/CMV2,应用脂质体将重组pRC/CMV2-hNK4质粒转入胰腺癌SW1990细胞中,采用逆转录聚合酶链反应(RT-PCR)及免疫印迹(Westernblot)分别检测转染的肿瘤细胞中NK4mRNA和蛋白的表达并筛选出高表达的细胞克隆。采用Transwall小室和Matrigel侵袭小室测定转染NK4基因对胰腺癌细胞运动和侵袭的影响。结果:转导NK4基因的SW1990细胞可表达并分泌NK4,RT-PCR扩增出预期的453bp片段,Westernblot显示有50000的NK4蛋白,转染NK4基因对胰腺癌细胞SW1990的生长无抑制作用(P>0.05),但可显著抑制HGF或成纤维细胞所诱导胰腺癌细胞的运动和侵袭(P<0.01),而转染空载体则无此作用(P>0.05)。结论:转染NK4基因可抑制胰腺癌细胞的运动和侵袭,在胰腺癌抗转移治疗中可能具有重要作用。 BACKGROUND &OBJECTIVE: Hepatocyte growth factor (HGF) plays an imp ortant role in the regulation of migration, invasion,and angiogenesis of cancer via the activation of its receptor, c-Met. NK4 is not only an antagonist of HGF but also an angiogenesis inhibitor. The blockade of HGF/c-Met signal pathway a nd tumor angiogenesis may be a new strategy for cancer treatment. This study was designed to construct eukaryotic expressing vector of NK4 gene, transfer it int o human pancreatic cancer cell line SW1990, and observe the effect of transfecte d NK4 gene on the biological behaviors of SW1990 cells,and its expression in SW1 990 cells. METHODS: The recombinant of pcDNA3/hNK4 plasmid was digested by restr ictive enzyme,NK4 gene was cloned into a high effective eukaryotic expressing ve ctor pRC/CMV2, and the recombinant of pRC/CMV2-hNK4 plasmid was transiently int roduced into SW1990 cells by lipofectamine. Reverse transcriptase-polymerase ch ain reaction (RT-PCR),and Western blot were used to detect the expression of NK 4 at mRNA,and protein levels,respectively. Migration, and invasion capabilities of the transfected cells were evaluated by Transwall chamber, and Matrigel invas ion chamber, respectively. RESULTS: Expressions of NK4 gene after lipofectamine mediated transfection were observed in SW1990 cells, expected fragment of 453 bp has been amplified by RT-PCR,and Western blot analysis showed positive express ion of NK4 protein (50 KDa). NK4 gene had no inhibitory effect on the growth of SW1990 cells (2.2×105 vs 2.5×105, P >0.05), while it had significantly suppres sive effect on the migration and invasion of SW1990 cells driven by HGF or fibro blasts (P< 0.01). CONCLUSION: NK4 gene transfection may inhibit spreading and in vasion of pancreatic cancer cells, which would play an important role in the ant i-metastasis therapy for pancreatic cancer.
出处 《癌症》 SCIE CAS CSCD 北大核心 2004年第10期1134-1138,共5页 Chinese Journal of Cancer
关键词 NK4基因 转染 胰腺癌细胞 HGF 侵袭 细胞生物学特性 运动 真核细胞表达载体 质粒 细胞克隆 NK4 gene Lipofectamine Gene therapy Pancreatic neoplasms
  • 相关文献

参考文献12

  • 1Date K, Matsu moto K, Shimura H, et al. HGF/NK4 is a specific antagonist for pleiotrophic actions of hepatocyte growth factor [J].FEBS Lett, 1997, 420:1-6.
  • 2Parr C, Jiang WG. Hepatocyte growth factor activators, inhibitors,and antagonists and their implication in cancer intervention [J] .Histol Histopathol, 2001, 16:251-268.
  • 3el-Kamar FG, Grossbard ML, Kozuch PS. Metastatic pancreatic cancer: emerging strategies in chemotherapy and palliative care [J]. Oncologist, 2003, 8(1): 18-34.
  • 4van der Voort R, Taher TE, Derksen PW, et al. The hepatocyte growth factor/Met pathway in development, tumorigenesis, and B-cell differentiation [J]. Adv Cancer Res, 2000, 79:39-90.
  • 5Ebert M, Yokoyama M, Friess H, et al. Coexpression of the c-met proto-oncogene and hepatocyte growth factor in human pancreatic cancer [J]. Cancer Res, 1994, 54:5775-5778.
  • 6Di Renzo MF, Poulsom R, Olivero M, et al. Expression of the Met/hepatocyte growth factor receptor in human pancrearic cancer [J]. Cancer Res, 1995,55: 1129-1136.
  • 7Paciucci R, Vial MR, Adell T, et al. Activation of the urokinase plasminogen activator/urokinase plasminogen activator receptor system and redistribution of E-cadherin are associated with hepatocyte growth factor-induced motility of pancreas tumor cells overexpressing Met [J]. Am J Pathol, 1998, 153:201-212.
  • 8Otte JM, Kiehne K, Schmitz F, et al. C-met protooncogene expression and its regulation by cytokines in the regenerating pancreas and in pancreatic cancer cells [J]. Scand J Gastroenterol,2000, 35(1): 90-95.
  • 9Matsumoto K, Nakamura T. NK4(HGFt/angiogenesis-antagonisinhibitor) in cancer biology and therapeutics [J]. Cancer Sci,2003, 94(4): 321-327.
  • 10赖人旭.肝细胞生长因子拮抗剂NK4在肿瘤中的研究进展[J].国外医学(肿瘤学分册),2002,29(2):101-104. 被引量:4

二级参考文献38

  • 1van der Voort R, Taher TE, Derksen PW, et al. The hepatocyte growth factor/Met pathway in development, tumorigenesis, and Bcell differentiation[J]. Adv Cancer Res, 2000,79(1) : 39-90.
  • 2Liu Y. The human hepatocyte growth factor receptor gene: eomplete structural organization and promoter characterization [J]. Gene, 1998, 215(1) : 159-169.
  • 3Weidner KM,Di Cesare S, Sachs M, et al. Interaction between Gabl and the c-Met receptor tyrosine kinase is responsible for epithelial morphogenesis[J]. Nature, 1996,384(6605) : 173-176.
  • 4To CT, Tsao MS. The roles of hepatocyte growth factor/scatter factor and met receptor in human cancers[J]. Oncol Rep, 1998,5 (5) : 1013-1024.
  • 5Ivan M, Bond JA, Prat M, et al. Activated ras and ret oncogenes induce over-expression of c-met (hepatocyte growth factor receptor) in human thyroid epithelial cells[J]. Oncngene, 1997, 14 (20) : 2417-2423.
  • 6Webb CP, Taylor GA, Jeffers M, et al. Evidence for a role of MetHGF/SF during Ras-mediated tumorigenesis/metastasis [J]. Oncogene, 1998,17(16) :2019-2025.
  • 7Schmidt L, Duh FM, Chen E, et al. Germline and somatic mutations in the tyrosine kinase domain of the MET proto-oncogene in papillary renal carcinomas[J]. Nat Genet, 1997,16(1) : 68-73.
  • 8Jeffers M, Schmidt L, Nakaigawa N, et al. Activating mutations for the met tyrosine kinase receptor in human cancer[J]. Proc Natl Acad Sci USA, 1997,94(21) : 11445-11450.
  • 9Toi M, Taniguchi T, Ueno T, et al. Significance of circulating hepatocyte growth factor level as a prognostic indicator in primary breast cancer[J]. Clin Cancer Res, 1998,4(3) :659-664.
  • 10Nagy J, Curry GW, Hillan K J, et al. Hepatocyte growth factor/scatter factor expression and c-met in primary breast cancer[J]. Surg Oncol, 1996,5(1) : 15-21.

共引文献3

同被引文献43

  • 1李静喆,张辉,唐梅,张仲良,张永清,李其棠.以HGF/c-Met为靶点治疗结肠癌及其与MEK2/ERK信号传导通路的关系[J].中华实验外科杂志,2005,22(4):504-504. 被引量:9
  • 2冯仟佳,杨涛,解军,牛勃,韩锐,陈修贵,梁冬明.不同浓度NK4对多种肿瘤细胞侵袭能力的影响[J].中国药物与临床,2005,5(11):834-836. 被引量:1
  • 3STUART K A, RIORDAN S M, LIDDER S, et al. Hepatocyte growth factor/scatter factor-induced intracellular signaling[J]. Int J Exp Pathol, 2000,81: 17-30.
  • 4DATE K, MATSUMOTO K, SHIMURA H, et al. HGF/NK4 is a specific antagonist for pleiotrophic actions of hepatocye growth factor[J]. FEBS Lett, 1997,420:1-6.
  • 5DATE K, MATSUMOTO K, NAKAMURA T, et al. Inhibition of tumor growth and invasion by a four- Kringle antagonist ( HGF/NK4 ) for hepatocyte growth factor[J]. Oncogene, 1998,17 : 3045 - 3054.
  • 6PARR C, HISCOX S,NAKAMURA T,et al. NK4,a new HGF/SF variant, is an antagonist to the influence of HGF/SF on the motility and invasion of colon cancer cells[J]. Int J Cancer,2000,85:563-570.
  • 7SON G, HIRANO T, SEKI E ,et al. Blockage of HGF/c-Met system by gene therapy (adenovirus-mediated NK4 gene) suppresses hepatocellular careinomainmice[J]. J Hepatol,2006,45:688-695.
  • 8HISCOX S, PARR C, MATSUMOT K,et al. NK4, a HGF/SF variant, inhibits in vitro invasiveness of breast cancer cells [J]. Breast Cancer Res Treat, 2000,59:245-254.
  • 9DATE K, MATSUMOTO K, KUBA K, et al. Inhibition of tumor growth and invasion by a four-kringle antagonist (HGF/NK4) for hepatocyte growth factor [J]. Oncogene, 1998,17 : 3045 - 3054.
  • 10PARR C, HISCOX S, CAI J, et al. Inhibition of HGF/SF and fibroblasts induced motility and invasion of colorectal cancer cells by NK4, a new HGF antagonist[J]. Int J Cancer, 2000,85:563-570.

引证文献3

二级引证文献5

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部