Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (27): 4993-4996.doi: 10.3969/j.issn.1673-8225.2010.27.012

Previous Articles     Next Articles

Effects of Notch1 gene silencing in mouse embryonic stem cells on glycogen synthase kinase-3 beta and cyclin-dependent kinase 5 expression

Zhang Qiu-ying 1,2 , Duan Ping1, Han Xue-fei1, Xu Yan1, Xing Ying1, Yan Wen-hai1   

  1. 1 Stem Cell Research Center, Medical College, Zhengzhou University, Zhengzhou 
    450052, Henan Province, China;2 Zhengzhou Shuqing Medical  School, Zhengzhou  450064, Henan Province, China
  • Online:2010-07-02 Published:2010-07-02
  • Contact: Yan Wen-hai, Doctor, Professor, Stem Cell Research Center, Medical College, Zhengzhou University, Zhengzhou 450052, Henan Province, China yanwh@zzu.edu.cn
  • About author:Zhang Qiu-ying, Master, Stem Cell Research Center, Medical College, Zhengzhou University, Zhengzhou 450052, Henan Province, China; Zhengzhou Shuqing Medical School, Zhengzhou 450064, Henan Province, China zqy497741121@126.com
  • Supported by:

    the Outstanding Youth Foundation of Henan Province, No.  94100510025*

PDF

372

Abstract:

BACKGROUND: Glycogen synthase kinase-3β (Gsk-3β) and cyclin-dependent kinase 5 (Cdk5) are phosphorylated protein kinase, and participate in differentiation and development of embryonic neuronal cells, but the precise mechanism remains unclear.
OBJECTIVE: To detect the changes in genetic expression of Hes-1, PS1, Gsk-3β and Cdk5 after Notch1 gene silencing in embryonic stem cells (ESCs), thus to explore Notch1 signaling accommodative action on Gsk-3β and Cdk5 and its potential impact on early neurogenesis.     
METHODS: Lipofectamine 2000 was used to transfer Notch1 small interfering RNA (siRNA) eukaryotic expression vector into mouse ESCs. Morphologic changes of cells before and after transfection were observed under inverted microscope. ESCs were collected at 48 hours after transfection. Genetic expressions of Notch1, Hes-1, PS1, Gsk-3β and Cdk5 in these cells are detected by RT-PCR. The intergroup differences were analyzed using one-way variance analysis.
RESULTS AND CONCLUSION:  ①Relative expression of Notch1, Hes-1, Gsk-3β and Cdk5 in pSINsi-U6-Notch1 group was obviously lower than blank plasmid and normal control groups ( P <0.05). There were no significant differences between blank plasmid and normal control groups (P > 0.05, n=5). ②Under an inverted microscope, exogenous plasmid did not impact ESCs cloning growth, but the proliferation quantity of cells in pSINsi-U6-Notch1 group was decreased compared with blank plasmid and normal control groups. ③No significant differences in genetic expression of PS1 were detected among the three groups (P > 0.05). Decreased Notch1 gene expression in ESCs down-regulates gene expression of Gsk-3β and Cdk5.

CLC Number: