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IGF-FOXO3a pathway regulates self-renewal of liver cancer stem cells in vitro



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IGF-FOXO3a pathway regulates self-renewal of liver cancer stem cells in vitro


LI ZhifenSHEN JunjieSHAN JuanjuanCHEN JunQIAN Cheng

College of Life Science, Zhejiang Sci-Tech University, Hangzhou, Zhejiang Province, 310018; Biological Therapy Center, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China


liver cancer stem cells FOXO3a insulin-like growth factor colony formation ability sphere formation ability


Objective      To investigate the role of FOXO3a in the regulation of self-renewal capacity in liver cancer stem cells (LCSCs) and study the related mechanisms. Methods     Flow cytometry was used to obtain LCSCs,and then the expression of FOXO3a in the attained cells was detected by real time fluorescent quantitative PCR (qRT-PCR). After the activity of insulin-like growth factor(IGF) was changed with IGF inhibitor or activator, the expression of FOXO3a was measured again with Western blotting and qRT-PCR. The lentiviral vectors expressing or interfering FOXO3a were used to infect the cells, and Western blot analysis was used to detect the efficiencies of overexpression and interference. Cell colony formation assay and sphere formation assay were used to test the self-renewal in the cells infected with the lentivirus that expressing or interfering FOXO3a, and the cells treated with IGF activators meanwhile over-expressing FOXO3a. Results     The expression of FOXO3a was significantly lower in the LCSCs than the liver cancer cells (P<0.01), and it was negatively regulated by IGF. The cell models of stable overexpression or interference of FOXO3a were established successfully. And FOXO3a over-expression resulted in decreases in the colony formation and sphere formation,while FOXO3a interference led to improvement in the formations. Activation of IGF signal, combined with FOXO3a overexpression also induced decreased formations. Conclusion     FOXO3a exerts a negative effect on the self-renewal in LCSCs, and can be negatively regulated by IGF signaling. It mediates the regulation of IGF signaling on the selfrenewal in LCSCs.


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Last Update: 2017-08-23