Functional analysis of Tcl1 using Tcl1-deficient mouse embryonic stem cells.
Functional analysis of Tcl1 using Tcl1-deficient mouse embryonic stem cells.
复制标题
DOI：
10.1371/journal.pone.0071645
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Miyazaki J
中科院分区：
文献类型：
--
作者：
Miyazaki T;Miyazaki S;Ashida M;Tanaka T;Tashiro F;Miyazaki J
Tcl 1在胚胎干（ES）细胞中高度表达，但其表达在分化后迅速降低。为了评估Tcl 1在ES细胞中的作用，我们产生了Tcl 1缺陷和过表达的小鼠ES细胞系。我们发现，Tcl 1是既不必要，也不足以维持未分化状态。据报道，Tcl 1激活Akt并增强细胞增殖。我们发现Tcl 1的表达水平与ES细胞的增殖率呈正相关，与凋亡呈负相关，但不影响Akt的磷酸化。另一方面，β-catenin的磷酸化水平随着Tcl 1的过表达而降低。我们使用TOPflash报告基因测定法测量了β-catenin活性，发现野生型ES细胞具有低活性，其中Tcl 1过表达增强了1.8倍。当经典Wnt信号传导被β-连环蛋白稳定化激活时，据报道它有助于维持ES细胞处于未分化状态。然后，我们进行了Tcl 1缺陷和表达ES细胞之间的DNA微阵列分析。结果表明，Tcl 1表达下调一组不同的基因，包括Ndp 52，其表达是非常高的囊胚，但减少在原始外胚层。基于这些结果，我们讨论了Tcl 1在ES细胞中的可能作用。
Tcl1 is highly expressed in embryonic stem (ES) cells, but its expression rapidly decreases following differentiation. To assess Tcl1’s roles in ES cells, we generated Tcl1-deficient and -overexpressing mouse ES cell lines. We found that Tcl1 was neither essential nor sufficient for maintaining the undifferentiated state. Tcl1 is reported to activate Akt and to enhance cell proliferation. We found that Tcl1 expression levels correlated positively with the proliferation rate and negatively with the apoptosis of ES cells, but did not affect Akt phosphorylation. On the other hand, the phosphorylation level of β-catenin decreased in response to Tcl1 overexpression. We measured the β-catenin activity using the TOPflash reporter assay, and found that wild-type ES cells had low activity, which Tcl1 overexpression enhanced 1.8-fold. When the canonical Wnt signaling is activated by β-catenin stabilization, it reportedly helps maintain ES cells in the undifferentiated state. We then performed DNA microarray analyses between the Tcl1-deficient and -expressing ES cells. The results revealed that Tcl1 expression downregulated a distinct group of genes, including Ndp52, whose expression is very high in blastocysts but reduced in the primitive ectoderm. Based on these results, we discuss the possible roles of Tcl1 in ES cells.
登录
查看更多内容
通过表达分析在胚胎干细胞中解剖OCT3/4调节的基因网络。
影响因子：
3.7
作者：
Matoba, Ryo;Niwa, Hitoshi;Masui, Shinji;Ohtsuka, Satoshi;Carter, Mark G.;Sharov, Alexei A.;Ko, Minoru S. H.
通讯作者：
Ko, Minoru S. H.
影响因子：
20.3
作者：
Kang, SM;Narducci, MG;Russo, G
通讯作者：
Russo, G
影响因子：
23.9
作者：
Hayashi K;de Sousa Lopes SMC;Tang F;Lao K;Surani MA
通讯作者：
Surani MA
影响因子：
23.9
作者：
Najm, Fadi J.;Chenoweth, Josh G.;Anderson, Philip D.;Nadeau, Joseph H.;Redline, Raymond W.;McKay, Ronald D. G.;Tesar, Paul J.
通讯作者：
Tesar, Paul J.
影响因子：
4.6
作者：
Lickert, H;Cox, B;Rossant, J
通讯作者：
Rossant, J