Connexin 43 is involved in the generation of human-induced pluripotent stem cells
Connexin 43 is involved in the generation of human-induced pluripotent stem cells
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Connexin 43 参与人类诱导多能干细胞的生成
DOI：
10.1093/hmg/ddt074
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发表时间：
2013-06-01
影响因子：
3.5
通讯作者：
Xiang, Andy Peng
中科院分区：
文献类型：
--
作者：
Ke, Qiong;Li, Li;Xiang, Andy Peng
虽然体细胞可以通过异位表达确定的转录因子成功地编程以产生多能干细胞，但重编程效率低并且重编程机制仍不清楚。以前的报道表明，几乎所有的人连接蛋白（CX）的异构体表达的人胚胎干细胞（hES）和间隙连接细胞间通讯（GJIC）是重要的ES细胞的生存和分化。然而，CX在人诱导多能干细胞（iPS）中的表达谱以及CX在重编程回iPS细胞的过程中的作用仍然未知。在这里，我们确定了人胚胎成纤维细胞（hEF）和hEF衍生的iPS细胞中大多数形式的CX的表达水平。刮片加载/染料转移测定显示，人iPS细胞含有功能性间隙连接（GJ），其可受到GJ功能的药理学抑制剂的影响。我们发现CX43是重编程后最显著上调的CX。最重要的是，CX43的异位表达显著提高了重编程效率，而shRNA介导的内源性CX43表达的敲低大大降低了效率。此外，我们发现，CX43过表达或敲低影响E-CADHERIN的表达，E-CADHERIN是间充质-上皮转化（MET）的标志物，在重编程过程中。总之，我们的数据表明，CX43的表达是重要的重编程，并可能介导的MET是与收购的多能性。
Although somatic cells can be successfully programmed to create pluripotent stem cells by ectopically expressing defined transcriptional factors, reprogramming efficiency is low and the reprogramming mechanism remains unclear. Previous reports have shown that almost all human connexin (CX) isoforms are expressed by human embryonic stem (hES) cells and that gap junctional intercellular communication (GJIC) is important for ES cell survival and differentiation. However, the CX expression profiles in human induced pluripotent stem (iPS) cells and the role of CXs in the process of reprogramming back to iPS cells remains unknown. Here, we determined the expression levels of most forms of CX in human embryonic fibroblasts (hEFs) and in the hEF-derived iPS cells. A scrape loading/dye transfer assay showed that human iPS cells contained functional gap junctions (GJs) that could be affected by pharmacological inhibitors of GJ function. We found that CX43 was the most dramatically upregulated CX following reprogramming. Most importantly, the ectopic expression of CX43 significantly enhanced the reprogramming efficiency, whereas shRNA-mediated knockdown of endogenous CX43 expression greatly reduced the efficiency. In addition, we found that CX43 overexpression or knockdown affected the expression of E-CADHERIN, a marker of the mesenchymal-to-epithelial transition (MET), during reprogramming. In conclusion, our data indicate that CX43 expression is important for reprogramming and may mediate the MET that is associated with the acquisition of pluripotency.