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首页> 外文期刊>Cellular Signalling >TGFβ and BMP-2 regulate epicardial cell invasion via TGFβR3 activation of the Par6/Smurf1/RhoA pathway
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TGFβ and BMP-2 regulate epicardial cell invasion via TGFβR3 activation of the Par6/Smurf1/RhoA pathway

机译:TGFβ和BMP-2通过TGFβR3激活Par6 / Smurf1 / RhoA途径调节心外膜细胞的侵袭

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摘要

Coronary vessel development requires transfer of mesothelial cells to the heart surface to form the epicardium where some cells subsequently undergo epithelial-mesenchymal transformation (EMT) and invade the subepicardial matrix. Tgfbr3 ~(-/-) mice die due to failed coronary vessel formation associated with decreased epicardial cell invasion but the mediators downstream of TGFβR3 are not well described. TGFβR3-dependent endocardial EMT stimulated by either TGFβ2 or BMP-2 requires activation of the Par6/Smurf1/RhoA 1pathway where Activin Receptor Like Kinase (ALK5) signals Par6 to act downstream of TGFβ to recruit Smurf1 to target RhoA for degradation to regulate apical-basal polarity and tight junction dissolution. Here we asked if this pathway was operant in epicardial cells and if TGFβR3 was required to access this pathway. Targeting of ALK5 in Tgfbr3 ~(+/+) cells inhibited loss of epithelial character and invasion. Overexpression of wild-type (wt) Par6, but not dominant negative (dn) Par6, induced EMT and invasion while targeting Par6 by siRNA inhibited EMT and invasion. Overexpression of Smurf1 and dnRhoA induced loss of epithelial character and invasion. Targeting of Smurf1 by siRNA or overexpression of constitutively active (ca) RhoA inhibited EMT and invasion. In Tgfbr3 ~(-/-) epicardial cells which have a decreased ability to invade collagen gels in response to TGFβ2, overexpression of wtPar6, Smurf1, or dnRhoA had a diminished ability to induce invasion. Overexpression of TGFβR3 in Tgfbr3 ~(-/-) cells, followed by siRNA targeting of Par6 or Smurf1, diminished the ability of TGFβR3 to rescue invasion demonstrating that the Par6/Smurf1/RhoA pathway is activated downstream of TGFβR3 in epicardial cells.
机译:冠状动脉发育需要将间皮细胞转移到心脏表面以形成心外膜,其中一些细胞随后会经历上皮-间质转化(EMT)并侵入心包膜下基质。 Tgfbr3〜(-/-)小鼠死于冠状血管形成失败,与心外膜细胞侵袭减少有关,但尚未充分描述TGFβR3下游的介质。由TGFβ2或BMP-2刺激的依赖TGFβR3的心内膜EMT需要激活Par6 / Smurf1 / RhoA 1途径,其中激活素受体样激酶(ALK5)发出信号,Par6参与TGFβ的下游以募集Smurf1来靶向RhoA,从而降解以调节根尖。基础极性和紧密连接溶解。在这里,我们询问在心外膜细胞中该途径是否有效,以及是否需要TGFβR3才能进入该途径。在Tgfbr3〜(+ / +)细胞中靶向ALK5可抑制上皮特性的丧失和侵袭。野生型(wt)Par6的过表达,而不是显性阴性(dn)Par6的过表达,诱导EMT和侵袭,而siRNA靶向Par6则抑制了EMT和侵袭。 Smurf1和dnRhoA的过度表达引起上皮性状的丧失和侵袭。 siRNA靶向Smurf1或组成型活性(ca)RhoA的过度表达抑制EMT和侵袭。在Tgfbr3〜(-/-)心外膜细胞中,其对TGFβ2的侵袭能力降低,其中wtPar6,Smurf1或dnRhoA的过度表达诱导侵袭的能力降低。 Tgfbr3〜(-/-)细胞中TGFβR3的过表达,然后是Par6或Smurf1的siRNA靶向,降低了TGFβR3挽救侵袭的能力,表明Par6 / Smurf1 / RhoA途径在心外膜细胞中TGFβR3的下游被激活。

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