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EGFR过表达对原代肝细胞AKT的激活作用

EGFR Overexpression Activates the AKT Signaling in Primary Hepatocytes
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摘要 目的构建含表皮生长因子受体EGFR基因的重组腺病毒载体,并观察所构建腺病毒介导的EGFR过表达对其下游的AKT通路的激活。方法将EGFR的cDNA片段克隆至pshuffle-CMV载体,将该载体与pAdEasy质粒进行细菌内同源重组从而获得重组腺病毒载体pAd-EGFR,之后在Ad293细胞中进行包装以及扩增,并对病毒效价进行检测;最后利用所制备的腺病毒Ad-EGFR感染小鼠原代肝细胞,利用蛋白印迹法Western blot检测EGFR在原代肝细胞中的表达及其对信号分子AKT的活化作用。结果成功制备具有生物学活性的EGFR重组腺病毒,并观察到该重组腺病毒在Huh-7细胞介导EGFR的过表达以及在原代肝细胞中对EGFR下游PI3K-AKT通路的激活作用。结论所构建的EGFR重组腺病毒载体可以介导细胞中EGFR的过表达并激活PI3K-AKT信号通路,为今后研究EGFR的相关生物学功能奠定了基础。 Objective To prepare EGFR recombinant adenovirus, and examine the activation of PI3K - AKT pathway in primary hepatoeytes by EGFR overexpression. Methods Mouse EGFR cDNA was first cloned into the shuttle vector pShuttle - CMV, then re- combined with pAdEasy - 1 plasmid in E. coli strain BJ5183 to obtain the pAd - EGFR vector, which was then transfected into 293A cells for viral package. Huh - 7 cells and mouse primary hepatocytes were infected with adenovirus Ad - EGFR and examined EGFR expression of and AKT phosphorylation by Western blotting analysis. Results We successfully prepared EGFR recombinant adenoviruses, and found that EGFR overexpression activates PI3 K - AKT pathway in primary hepatocytes. Conclusion The established EGFR adenovirus vector may provide a useful tool in investigating the role of EGFR in liver biology.
出处 《医学研究杂志》 2013年第12期19-22,共4页 Journal of Medical Research
基金 国家自然科学基金资助项目(81130084)
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参考文献11

  • 1Yarden Y. The EGFR family and its ligands in human cancer.Signalling mechanisms and therapeutic opportunities[J].{H}EUROPEAN JOURNAL OF CANCER,2001,(Suppl 4):S3-S8.
  • 2Hayase Y,Higashiyama S,Sasahara M. Expression of heparin-binding epidermal growth factor-like growth factor in rat brain[J].{H}Brain Research,1998,(1-2):163-178.
  • 3Huang HS,Nagane M,Klingbeil CK. The enhanced tumorigenic activity of a mutant epidermal growth factor receptor common in human cancers is mediated by threshold levels of constitutive tyrosine phosphorylation and unattenuated signaling[J].{H}Journal of Biological Chemistry,1997,(05):2927-2935.
  • 4Ai Z,Wang J,Wang Y. Overexpressed epidermal growth factor receptor (EGFR)-induced progestin insensitivity in human endometrial carcinoma cells by the EGFR/mitogen-activated protein kinase signaling pathway[J].{H}CANCER,2010,(15):3603-3613.
  • 5Zhang Y,Xie Z,Zhou L. The zinc finger protein ZBTB20 regulates transcription of fructose-1,6-bisphosphatase 1 and beta cell function in mice[J].{H}GASTROENTEROLOGY,2012,(07):1571-1580.
  • 6Lambotte L,Saliez A,Triest S. Effect of sialoadenectomy and epidermal growth factor administration on liver regeneration after partial hepatectomy[J].{H}HEPATOLOGY,1997,(03):607-612.
  • 7Natarajan A,Wagner B,Sibilia M. The EGF receptor is required for efficient liver regeneration[J].{H}Proceedings of the National Academy of Sciences(USA),2007,(43):17081-17086.doi:10.1073/pnas.0704126104.
  • 8Lupberger J,Zeisel MB,Xiao F. EGFR and EphA2 are host factors for hepatitis C virus entry and possible targets for antiviral therapy[J].{H}Nature Medicine,2011,(05):589-595.
  • 9da Cunha Santos G,Shepherd FA,Tsao MS. EGFR mutations and lung cancer[J].Annu Rev Pathol,2011.49-69.
  • 10Chettouh H,Fartoux L,Aoudjehane L. Mitogenic insulin receptor-A is overexpressed in human hepatocellular carcinoma due to EGFR-mediated dysregulation of RNA splicing factors[J].{H}CANCER RESEARCH,2013,(13):3974-3986.

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