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RhoC基因真核表达载体的构建及其表达

Construction of RhoC gene vector and its expression in eucaryotic cells
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摘要 目的　 为探讨RhoC基因的功能以及其对肝癌的侵袭转移的影响提供实验模型。方法　用限制性内切酶酶切RhoC基因 ,定向克隆到 pcDNA 3 .1上。利用脂质体将重组载体 (pcDNA 3 .1 RhoC)和空载体 (pcDNA 3 .1 )转染入HEPG2细胞 ,潮霉素选择培养 ,RT PCR及免疫组化鉴定其稳定表达。 结果　构建的真核表达载体 pcDNA 3 .1 RhoC在HEPG2细胞中有稳定表达。 结论　本实验为深入研究RhoC基因的功能以及其对肝癌的侵袭转移的影响提供了理想的体外实验模型。 Objective To establish an experimental model for exploring the role of RhoC gene in the invasiveness and metastasis of hepatocellular carcinoma.Method The RhoC gene was digested with restricted enzyme Hind III and XbaI,and direct cloned to pcDNA3.1.The recombinant vector (pcDNA3.1 RhoC) and the vector alone (pcDNA3.1) were transfected into HEPG2 cells with LIPFECTAMINETMReagent.After selected with hygromycin,resistant cloneies was obtained.The transcription and translation of RhoC gene were analysed with the reverse transcription PCR and immunohistochemical stain.Results The recombinant vectort (pc DNA3.1 Rhoc) express steadily in HerpG2 cells.Conclusions The modified tumor cells(HEPG2 RhoC) could be used to study the effect of RhoC protein on the invasiveness and metastasis of hepatocellular carcinoma.
作者 郑岩松 吕新生 林永堃
机构地区 福建医科大学附属第一医院普外科 中南大学湘雅医院普外科
出处 《中国普通外科杂志》 CAS CSCD 2002年第12期737-740,共4页 China Journal of General Surgery
基金 福建省政府科研基金资助 ( 99J- 0 0 6 ) 福建省卫生厅青年科研基金资助。 ( 2 0 0 2 - 0 1- 0 4 )
关键词 RHOC基因 真核表达 载体 构建 肝肿瘤 基因表达 GENE RhoC GENE EXPRESSION REGULATION, NEOPLASTIC LIVER NEOPLASMS/genet
分类号 R735.7 [医药卫生—肿瘤]
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参考文献10

  • 1Madaule P,Axel R,et al.A novel ras-related gene family[J]. Cell,1985,41(1):31-40.
  • 2Chardin P,Madaule P,Tavitian A,et al. Coding sequence of human rho cDNAs clone 6 and clone 9[J]. Nucleic Acid Res, 1988,16(6): 2717.
  • 3Fagan K,Oliveira L,Pittler S.Sequence of rho small GTP-binding protein cDNAs from human retina and identification of novel 5-prime end cloning artifacts[J].Exp Eye Res, 1994,59(2): 235-237.
  • 4萨姆布鲁克 J,弗里奇 EF,曼尼阿蒂斯 T.分子克隆:实验指南[M].第2版.金冬雁,黎孟枫译.北京:科学出版社,1992.316-472.
  • 5Maekawa M,Ishizaki T,Boku S , et al. Signaling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase[J]. Science, 1999,285(5429):895-898.
  • 6Clark E, Golub T,Lander E,et al. Genomic analysis of metastasis reveals an essential role for RhoC[J]. Nature,2000,406(6795):532-535.
  • 7Kleer CG, van Golen KL, Zhang Y,et al.Characterization of RhoC expression in benign and malignant breast disease :A potential new marker for small breast carcinomas with metastatic ability[J].Am J Pathol, 2002, 160(2):579-584.
  • 8石铮,郑岩松,曾金华,刘景丰,林旭.RhoC基因及蛋白在胃癌肝转移中的表达及意义[J].中华实验外科杂志,2002,19(4):310-311. 被引量：23
  • 9Takamura M, Sakamoto M,Genda T, et al.Inhibition of intrahepatic metastasis of human hepatocellular carcinoma by Rho-associated protein kinase inhibitor Y-27632[J].Hepatology, 2001,33(3):577-581.
  • 10郑岩松,吕新生,林永堃,石铮.RhoC基因在原发性肝癌中的表达及其意义[J].中国普通外科杂志,2002,11(9):528-530. 被引量：7

二级参考文献11

  • 1[1]Madaule P, Axel R. A novel ras-related gene family[J]. Cell,1985,41(1):31-40.
  • 2[2]Chardin P,Madaule P,Tavitian A,et al. Coding sequence of human Rho cDNAs clone 6 and clone 9[J]. Nucleic Acid Res, 1988,16(): 2717.
  • 3[3]Clark E, Golub T, Lander,E, et al. Genomic analysis of metastasis reveals an essential role for RhoC[J]. Nature, 2000,406(): 532-535.
  • 4[4]Kleer CG, van Golen KL, Zhang Y,et al.Characterization of RhoC expression in benign and malignant breast disease : A potential new marker for small breast carcinomas with metastatic ability[J]. Am J Pathol, 2002, 160(2):579-584.
  • 5[5]Takamura M, Sakamoto M, Genda T, et al. Inhibition of intrahepatic metastasis of human hepatocellular carcinoma by Rho-associated protein kinase inhibitor Y-27 632[J]. Hepatology, 2001,33(3):577-581.
  • 6[6]Genda T,Sakamoto M,Ichida T,et al.Cell motility mediated by Rho and Rho-associated protein kinase plays a critical role in intrahepatic metastasis in human hepatocellular carcinoma[J]. Hepatology, 1999,30():1027-1036.
  • 7[7]Hiroshi O,Ssiji S,Tatsushi K,et al.Genome-wide analysis of gene expression in human hepatocellular carcinomas using cDNA microssary:Identification of genes involved in viral carcinogenesis and tumor progression[J].Cancer Res,2001, 61():2129-2137.
  • 8Kleer CG,Van Golen KL,Zhang Y,et al.Characterization of RhoC expression in benign and malignant breast disease: a potential new marker for small breast carcinomas with metastatic ability[].American Journal of Pathology.2002
  • 9Clark EA,Golub TR,Lander ES,et al.Genomic analysis of metastasis reveals an essential role for RhoC[].Nature.2000
  • 10Takamura M,Sakamoto M,Genda T,et al.Inhibition of intrahepatic metastasis of human hepatocellular carcinoma by Rho-associated protein kinase inhibitor Y-27632[].Hepatology.2001

共引文献27

中国普通外科杂志
2002年 第12期

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