VEGF-mediated PI3K class IA and PKC signaling in cardiomyogenesis and vasculogenesis of mouse embryonic stem cells
VEGF-mediated PI3K class IA and PKC signaling in cardiomyogenesis and vasculogenesis of mouse embryonic stem cells
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DOI：
10.1242/jcs.077594
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发表时间：
2011-06-01
影响因子：
4
通讯作者：
Wartenberg, Maria
中科院分区：
文献类型：
--
作者：
Bekhite, Mohamed M.;Finkensieper, Andreas;Wartenberg, Maria
在小鼠ES细胞和ES细胞衍生的Flk-1（+）心血管祖细胞中研究了VEGF、磷酸肌醇3-激酶（PI 3 K）和蛋白激酶C（PKC）调节的心脏和血管分化中的信号传导。通过渥曼青霉素和LY 294002抑制PI 3 K，使用短发夹RNA（shRNA）破坏PI 3 K催化亚基p110 α和p110 δ，或用化合物15 e抑制p110 α和用IC-87114抑制p110 δ，损害心脏和血管分化。相比之下，TGX-221，一种p110 β的抑制剂，和p110 β的shRNA敲低没有显着的效果。PKC家族的拮抗剂，即双吲哚基马来酰亚胺-1（BIM-1）、G 6976（靶向PKC α/β II）和rottlerin（靶向PKC δ）消除血管发生，但不消除心肌发生。Akt的抑制钝化心脏以及血管分化。VEGF诱导PKC α/β II和PKC δ磷酸化，但不诱导PKC zeta磷酸化。这被PI 3 K抑制剂和VEGFR-2拮抗剂SU 5614消除。此外，Akt和磷酸肌醇依赖性激酶-1（PDK 1）的磷酸化在抑制PI 3 K时减弱，但在BIM-1抑制PKC时不减弱，表明VEGF激活Akt和PDK 1需要PI 3 K而不是PKC。总之，我们证明了PI 3 K催化亚基p110 α和p110 δ是ES细胞心血管发生的中心。PI 3 K下游的Akt参与心肌发生和血管发生，而PKC仅参与血管发生。
VEGF-, phosphoinositide 3-kinase (PI3K)- and protein kinase C (PKC)-regulated signaling in cardiac and vascular differentiation was investigated in mouse ES cells and in ES cell-derived Flk-1(+) cardiovascular progenitor cells. Inhibition of PI3K by wortmannin and LY294002, disruption of PI3K catalytic subunits p110 alpha and p110 delta using short hairpin RNA (shRNA), or inhibition of p110 alpha with compound 15e and of p110 delta with IC-87114 impaired cardiac and vascular differentiation. By contrast, TGX-221, an inhibitor of p110 beta, and shRNA knockdown of p110 beta were without significant effects. Antagonists of the PKC family, i.e. bisindolylmaleimide-1 (BIM-1), G 6976 (targeting PKC alpha/beta II) and rottlerin (targeting PKC delta) abolished vasculogenesis, but not cardiomyogenesis. Inhibition of Akt blunted cardiac as well as vascular differentiation. VEGF induced phosphorylation of PKC alpha/beta II and PKC delta but not PKC zeta. This was abolished by PI3K inhibitors and the VEGFR-2 antagonist SU5614. Furthermore, phosphorylation of Akt and phosphoinositide-dependent kinase-1 (PDK1) was blunted upon inhibition of PI3K, but not upon inhibition of PKC by BIM-1, suggesting that activation of Akt and PDK1 by VEGF required PI3K but not PKC. In summary, we demonstrate that PI3K catalytic subunits p110 alpha and p110 delta are central to cardiovasculogenesis of ES cells. Akt downstream of PI3K is involved in both cardiomyogenesis and vasculogenesis, whereas PKC is involved only in vasculogenesis.