[Suppression of ras mediated signaling attenuates hematopoietic differentiation of embryonic stem cells of mice in vitro].
To investigate the possible involvement of Ras signaling in the hematopoietic differentiation of embryonic stem cells (ES cells), ES cells were transfected with RasN17, the dominant-negative mutant of Ras. Western blot was used to test the effect of RasN17 expression on Erk1/2 and Akt phosphorylation, semi-quantitative RT-PCR was used to detect expression of gene related to hematopoiesis in differentiation of ES cells. The results showed that the expression of RasN17 in the ES cells remarkably downregulated the phosphorylation of Erk1/2 and Akt simultaneously. Moreover, the expression of several markers related with hematopoiesis including Runx1, SCL and beta-major globin, were significantly suppressed in the EB expressing RasN17, whereas the transcription of Flk1, a gene required earlier than SCL in development of hematopoietic and endothelial lineages, was not influenced. It is concluded that the activation of Ras is pivotal for in vitro hematopoietic differentiation of ES cells.
Wang
Xiao-Yan
XY
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
Liu
Bing
B
Yao
Hui-Yu
HY
Hou
Ning
N
Yang
Xiao
X
Yu
Xiao-Dan
XD
Mao
Ning
N
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology*; D011905:Genes, ras; D006410:Hematopoiesis / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000166:cytology; D051379:Mice; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D010766:Phosphorylation; D015398:Signal Transduction; D018631:ras Proteins / Q000502:physiology*
D048052:Mitogen-Activated Protein Kinase 3; D018631:ras Proteins