Chinese Journal of Tissue Engineering Research

Previous Articles     Next Articles

Pten gene expression and phosphoproteomic analysis of the downstream of PI3K/Akt/mTOR signaling in human embryonic stem cells

Zhou Rui-qing, Gong Yu-ping, Xing Hong-yun, Yang Xi   

  1. Department of Hematology and Research Lab of Hematology, West China Hospital, Sichuan University, Chengdu  610041, Sichuan Province, China
  • Received:2011-06-18 Revised:2011-07-09 Online:2011-12-03 Published:2011-12-03
  • Contact: Gong Yu-ping, Professor, Department of Hematology and Research Lab of Hematology, West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China gongyuping2010@yahoo.cn
  • About author:Zhou Rui-qing☆, Doctor, Department of Hematology and Research Lab of Hematology, West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China zrqhematology@foxmail.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30770912*; Social Welfare Project of Sichuan Province Bureau of Science and Technology, No. 2008SZ0017*; the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, No. 20071108-18-4*

PDF

954

Abstract:

BACKGROUND: Phosphoprteomic analyses of human embryonic stem cells (hESCs) in undifferentiated state and their differentiated derivatives have become a hotspot. The researchs suggest that phosphorylation events determine hESCs’ fate.
OBJECTIVE: To investigate the expression of Pten mRNA and the key proteins of PTEN/Akt/mTOR signaling in human embryonic stem cells in order to provide the basic research for stem cells expansion and differentiation.
METHODS: The hESCs were cocultured with the murine fetal fibroblasts to maintain undifferentiation and digested by collagenase for detection. The growth morphology of hESCs were observed, RT-PCR assay was used to determine Pten gene expression, and Western blot assay to detect the expression of p-PTEN, p-mTOR, p-P70S6K, p-4E-BP1 in hESCs.
RESULTS AND CONCLUSION: The expression level of Pten was higher in hESCs and feeder cells than in K562, while the expression level of the key proteins in hESCs was lower than K562, especially p-4E-BP1 in hESCs. Over-activation of the PI3K/Akt/mTOR signaling may accelerate the proliferation of hESCs and deduce the apoptosis, and provide more cells for differentiation and regeneration medicine.

CLC Number: