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Gab2对人肺癌细胞A549侵袭能力影响及其机制的探讨 被引量：7

Role and mechanism of Gab2 in invasion of human lung cancer A549 cells
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摘要 目的:探讨Grb2协同结合蛋白2(Grb2binding protein-2,Gab2)对人肺癌A549细胞体外迁移和侵袭的影响及其机制。方法:将质粒pGCsilencerU6/GFP/Neo-RNAi-Gab2#1、pGCsilencerU6/GFP/Neo-RNAi-Gab2#2和对照空载质粒转染到A549细胞中,建立Gab2低表达的siGab2/A549#1、siGab2/A549#2细胞和对照SCR/A549细胞。应用RT-PCR和蛋白质印迹法检测小分子干扰RNA(siRNA)干扰Gab2表达后的基因和蛋白表达水平。趋化运动实验检测细胞的迁移能力;体外侵袭实验检测细胞的侵袭能力;蛋白质印迹法检测siRNA干扰Gab2表达后,A549细胞在胰岛素样生长因子-1(insulinlike growth factor-1,IGF-1)刺激下基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、基质金属蛋白酶-9(matrix metal-loproteinase-9,MMP-9)的表达及磷酸化Akt(pAkt)、磷酸化人雷帕霉素靶蛋白(phosphorylated mammalian target of rapamy-cin,pmTOR)的活化情况。结果:siRNA干扰Gab2表达后,RT-PCR测定A549细胞Gab2mRNA相对表达量为1.340±0.009,Scr/A549细胞为1.201±0.074,siGab2#1/A549细胞为0.315±0.008,siGab2#2/A549细胞为0.289±0.007。蛋白印迹法检测A549细胞Gab2蛋白表达量为0.205±0.003,Scr/A549细胞为0.241±0.004,siGab2#1/A549细胞为0.128±0.002,siGab2#2/A549细胞为0.066±0.001。siGab2#2/A549细胞与A549细胞比较Gab2基因表达显著降低,t=168.032,P<0.001;Gab2蛋白表达也显著降低,t=64.352,P<0.001。siGab2#2/A549细胞的体外迁移和侵袭能力明显低于SCR/A549细胞,t值分别为5.101和10.812,P值分别为0.029和0.002。在IGF-1刺激下,siGab2/A549细胞的MMP-2、MMP-9蛋白的表达量不再有明显变化,t值分别为-2.051和-2.652,P值分别为0.054和0.064。siGab2/A549细胞中pAkt、pmTOR的活化也显著抑制。结论:Gab2可能通过调节Akt/mTOR信号通路,促进A549细胞的侵袭能力。 OBJECTIVE： To investigate the effect and the molecular mechanism of Gab2 on migration and invasion of human lung adenocarcinoma cell line A549. METHODS： The plasmid pGCsilencerU6/GFP/Neo RNAi-Gab2 35 1, pGCsi- lencerU6/GFP/Neo-RNAi-Gab2 35 2 and the empty vector were transfected into A549 cells respectively to establish Gab2-deficient A549 cells （siGab2/A549 35 1, siGab2/A549 ~ 2 cells） and the SCR/A549 cells （as the control cells）. RT-PCR and Western blot analyse were used to test the mRNA and protein expression levels of Gab2. The abilities of mi- gration and invasion were determined by chemotaxis assay and invasion assay. The IGF-1 induced expression of MMP-2, MMP-9 and the activation of pAkt,and pmTOR in Gab2-deficient A549 cells and SCR/A549 cells were detected by West- ern blot. The Gab2 mRNA expression of A549 cell, Scr/A549 cell, siGab2 # 1/A549 cell and siGab2 ~ 2/A549 cell were
作者 史立宏 陶蓉 王云甲 张秀荣 孙秀宁 周风华 罗荣城 张宝刚
机构地区 潍坊医学院山东省应用药理学重点实验室 南方医科大学南方医院肿瘤中心 潍坊医学院呼吸病学 潍坊医学院病原微生物学教研室 潍坊医学院病理学教研室
出处 《中华肿瘤防治杂志》 CAS 北大核心 2013年第16期1207-1211,共5页 Chinese Journal of Cancer Prevention and Treatment
基金 国家自然科学基金(81072068) 山东省中青年科学家科研奖励基金〔(博士基金)2010BSB14050〕 山东省教育厅高等学校优秀青年教师国内访问项目(2009) 潍坊市科学技术发展计划(201201230)
关键词 Gab2 肺肿瘤 A549细胞 侵袭 Gab2 lung cancer~ A549cell~ invasion
分类号 R734.2 [医药卫生—肿瘤]
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参考文献18

  • 1Adams SJ,Aydin IT, Celebi JT. GAB2--a scaffolding protein in cancer[J]. Mol Cancer Res, 2012,10 (10) : 1265-1270.
  • 2Wang Y,Sheng Q, Spillman MA,et al. Gab2 regulates the mi- gratory behaviors and E cadherin expression via activation of the PI3K pathway in ovarian cancer cells[J]. Oncogene, 2011,31 (20) :2512 2520.
  • 3Herrera Abreu MT, Hughes WE, Mele K,et al. Gab2 regulates cytoskeletal organization and migration of mammary epithelial cells by modulating RhoA activation[J]. Mol Biol Ce11,2011,22 (1):105 116.
  • 4张珊珊,柯越海.支架蛋白Gab2信号调控与乳腺癌[J].中国生物化学与分子生物学报,2011,27(4):300-304. 被引量：3
  • 5Xu XL,Wang X,Chen ZL,et al. Overexpression of Grb2 associ- ated binder 2 in human lung caneer[J]. Int J Biol Sci, 2011,7 (5) :496-504.
  • 6王小霞,康佳丽,聂妙玲,杨文娟.靶向RhoA shRNA对卵巢癌HO8910细胞体外侵袭能力影响的研究[J].中华肿瘤防治杂志,2012,19(11):818-821. 被引量：1
  • 7于涛,刘刚,祖玲玲,王伟强,李永文.Plumbagin对人高转移大细胞肺癌细胞系抑制作用的研究[J].中华肿瘤防治杂志,2012,19(4):268-271. 被引量：3
  • 8Simister PC, Feller SM. Order and disorder in large multi-site docking proteins of the Gab family-implications for signalling complex formation and inhibitor design strategies[J]. Mol Bio syst,2012,8(1) :33-46.
  • 9Gu H,Neel BG. The "Gab" in signal transduction[J]. Trends Cell Biol,2003,13(3) : 122-130.
  • 10H unzicker-Dunn ME, Lope:Biladeau B, Law NC,et al. PKA and GAB2 play central roles in theFSH signaling pathway to PI3K and AKT in ovarian granulosa cells[J]. Proc Natl Acad Sci U S A,2012,109(44) :2979 2988.

二级参考文献53

  • 1Ostman A, Hellberg C, Bohmer F D. Protein-tyrosine phosphatases and cancer[ J ]. Nat Rev Cancer,2006,6 (4) :307- 320.
  • 2Eulenfeld R, Schaper F. A new mechanism for the regulation of Gab1 recruitment to the plasma membrane [ J]. J Cell Sci,2009, 122(Pt 1) :55-64.
  • 3WohrIe F U, Daly R J, Brummer T. Function, regulation and pathological roles of the Gab/DOS docking proteins [ J ]. Cell Commun Signal, 2009,7:22.
  • 4Nishida K, Hirano T. The role of Gab family scaffolding adapter proteins in the signal transduction of cytokine and growth factor receptors [J]. Cancer Sci, 2003, 94(12):1029-1033.
  • 5Gu H, Neel B G. The "Gab" in signal transduction[J]. Trends Cell Biol, 2003,13(3) :122-130.
  • 6Maus M, Medgyesi D, Kovesdi D, et al. Grb2 associated binder 2 couples B-cell receptor to cell survival [ J]. Cell Signal,2009, 21 ( 2 ) : 220 -227.
  • 7Pan X L, Ren R J, Wang G, et al. The Gab2 in signal transduction and its potential ro|e in the pathogenesis of Alzheimer's disease [ J]. Neurosci Bull, 2010,26 (3) :241-246.
  • 8Franke T F. PI3K/Akt : getting it right matters [ J]. Oncogene, 2008,27(50) :6473-6488.
  • 9Zhang T T, Li H, Cheung S M, ,et al. Phosphoinositide 3- kinase-regulated adapters in lymphocyte activation [ J ]. Immunol Rev ,2009, 232( 1 ) :255-272.
  • 10Lock L S, Royal I, Naujokas M A, et al. Identification of an atypical Grb2 carboxyl-terminal SH3 domain binding site in Gab docking proteins reveals Grb2-dependent and-independent recruitment of Gabl to receptor tyrosine kinases [ J ]. J Biol Chem, 2000,275(40) :31536-31545.

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中华肿瘤防治杂志
2013年 第16期

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