JAK2/STAT3 directs cardiomyogenesis within murine embryonic stem cells in vitro
JAK2/STAT3 directs cardiomyogenesis within murine embryonic stem cells in vitro
复制标题
DOI：
10.1634/stemcells.2004-0293
复制
发表时间：
2005-04-01
期刊：
影响因子：
5.2
通讯作者：
Gallicano, GI
中科院分区：
文献类型：
--
作者：
Foshay, K;Rodriguez, G;Gallicano, GI
心脏是发育过程中最先形成的器官；然而，对控制心脏分化初始阶段的机制知之甚少。为了研究这一过程，我们使用蛋白激酶表达筛选，将非跳动胚胎干细胞（ES）与跳动胚胎干细胞衍生的心肌细胞进行比较。我们发现JAK2在跳动区域的蛋白质水平增加了70%。药理抑制JAK2或使用显性/阴性JAK2均可减少胚状体（EBs）内的跳动，而使用显性/阳性JAK2的功能分析结果可显著诱导跳动。更重要的是，JAK2的特异性靶点STAT3被显性/阴性STAT3抑制，导致跳动区域几乎完全丧失。逆转录聚合酶链反应和STAT3抑制EBs的Western分析导致一些心脏特异性基因缺乏表达，其中许多基因在其启动子内含有STAT3 dna结合区。综上所述，这些数据表明JAK2/STAT3通路对心肌形成的初始阶段至关重要。
The heart is the first organ to form during development; however, little is known about the mechanisms that control the initial stages of cardiac differentiation. To investigate this process, we used a protein kinase expression screen, in which nonbeating embryonic stem (ES) cells were compared with beating ES cell-derived cardiomyocytes. We found that JAK2 experienced a 70% increase in protein levels within beating areas. Inhibition of JAK2 pharmacologically or by using dominant/negative JAK2 both resulted in diminished beating within embryoid bodies (EBs), whereas gain of function analysis using dominant/positive JAK2 resulted in a significant induction of beating. More important, inhibition of STAT3, a specific target of JAK2, by dominant/negative STAT3 resulted in the virtual complete loss of beating areas. Reverse transcription-polymerase chain reaction and Western analysis of STAT3-inhibited EBs resulted in lack of expression of several cardiac-specific genes, many of which contain within their promoter STAT3 DNA-binding regions. Taken together, the data reveal that the JAK2/STAT3 pathway is essential for initial stages of cardiomyogenesis.