You Position: Home > Paper
Serum response factor participates in RhoA-induced endothelial cell F-actin rearrangements
( views:132, downloads:14 )
- Author:
- No author available
- Journal Title:
- ACTA PHYSIOLOGICA SINICA
- Issue:
- 3
- DOI:
- No doi available
- Key Word:
- 血清反应因子;RhoA;肌动蛋白;内皮细胞
Abstract： 为进一步阐明RhoA调控人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)肌动蛋白骨架重构的分子机制,用逆转录病毒感染并筛选出稳定表达持续活化型RhoA(Q63LRhoA)和主导抑制型RhoA(T19NRhoA)的HUVECs.应用免疫组化和Western blot方法分析去血清前后HUVECs血清反应因子(serum response factor,SRF)的表达及定位,Rhodamine-Phalloidine染色观察F-actin动态变化.结果显示,Q63LRhoA组细胞核中SRF表达增加,F-actin重排形成大量应力纤维;T19NRhoA组中SRF表达较弱,F-actin无明显改变,无应力纤维形成.去血清后,正常HUVECs(对照组)和感染细胞中SRF的表达均显著增加,但其亚细胞定位明显不同.对照组去血清培养3 d,SRF主要定位在细胞核,去血清培养5 d,SRF出核转位入细胞浆.Q63LRhoA组SRF发生核滞留,不随去血清培养时间延长发生出核转位现象.T19NRhoA组SRF的表达主要定位于细胞核周.对照组去血清培养3 d,F-actin表达增加,同时形成大量应力纤维,去血清培养5 d,细胞F-actin表达下调,应力纤维解聚.Q63LRhoA组F-actin重构持续发生并形成大量应力纤维,但不随去血清培养时间延长发生明显解聚.而T19NRhoA组F-actin表达不随去血清时间延长而增加.上述结果提示,RhoA介导HUVECs F-actin的重构与SRF的核转位现象密切相关.
- [1]王翔,蔡绍皙,罗向东,等.脂多糖直接损伤血管内皮细胞后内皮细胞膜脂的修饰[J].2001,(6).
- [2]Hautmann M,Mack CP,Somlyo AV,etc.Smooth muscle differentiation marker gene expression is regulated by RhoA-mediated actin polymerization.[J].2001,276(1).
- [3]M, Amano,K, Chihara,K, Kimura,etc.Formation of actin stress fibers and focal adhesions enhanced by Rho-kinase.[J].1997,275(5304).
- [4]Sotiropoulos A.,Posern G.,Treisman R..Mutant actins demonstrate a role for unpolymerized actin in control of transcription by serum response factor[J].2002,13(12).
- [5]Sandrine, Etienne-Manneville,Alan, Hall.Rho GTPases in cell biology.[J].2002,420(6916).
- [6]Marie Vandromme,Ned Lamb,Cecile Gauthier-Rouviere,etc.The serum response factor nuclear localization signal: general implications for cyclic AMP-dependent protein kinase activity in control of nuclear translocation[J].1995,15(1).
- [7]J, Rech,I, Barlat,J L, Veyrune,etc.Nuclear import of serum response factor (SRF) requires a short amino-terminal nuclear localization sequence and is independent of the casein kinase II phosphorylation site.[J].1994,107 ( Pt 11).
- [8]Ridley AJ.,Wojciak-Stothard B.,Potempa S.,etc.Rho and Rac but not Cdc42 regulate endothelial cell permeability[J].2001,114(7).
- [9]Turnage RH,Yin HL,Wright K,etc.Burn-activated neutrophils and tumor necrosis factor-alpha alter endothelial cell actin cytoskeleton and enhance monolayer permeability.[J].2000,128(2).
- [10]Schratt Gerhard,Berger Jurgen,Philippar Ulrike,etc.Serum response factor is crucial for actin cytoskeletal organization and focal adhesion assembly in embryonic stem cells.[J].2002,156(4).