[1]李志芬,沈俊杰,单娟娟,等.IGF-FOXO3a通路对肝癌干细胞自我更新能力的调控[J].第三军医大学学报,2017,39(16):1631-1636.
 LI Zhifen,SHEN Junjie,SHAN Juanjuan,et al.IGF-FOXO3a pathway regulates self-renewal of liver cancer stem cells in vitro[J].J Third Mil Med Univ,2017,39(16):1631-1636.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
39卷
期数:
2017年第16期
页码:
1631-1636
栏目:
基础医学
出版日期:
2017-08-30

文章信息/Info

Title:
IGF-FOXO3a pathway regulates self-renewal of liver cancer stem cells in vitro
作者:
李志芬沈俊杰单娟娟陈军钱程
浙江理工大学生命科学学院;第三军医大学西南医院生物治疗中心
Author(s):
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

关键词:
肝癌干细胞FOXO3a类胰岛素生长因子克隆形成能力成球生长能力
Keywords:
liver cancer stem cells FOXO3a insulin-like growth factor colony formation ability sphere formation ability
分类号:
R341;R730.23;R735.7
文献标志码:
A
摘要:

目的     探讨FOXO3a对肝癌干细胞(liver cancer stem cells, LCSCs)自我更新能力的调控作用以及其相关机制。方法     流式细胞仪分选获取肝癌干细胞,实时荧光定量PCR(real time fluorescent quantitative PCR,qRT-PCR)检测FOXO3a在肝癌干细胞中的表达情况;利用类胰岛素生长因子 (insulinlike growth factor, IGF)的抑制剂或激活剂改变IGF活性,Western blot和qRT-PCR检测FOXO3a表达的改变;包装、纯化表达和干扰FOXO3a的慢病毒并进行细胞的感染,Western blot检测其过表达和干扰效率;采用细胞克隆形成实验和细胞成球生长能力实验检测过表达或干扰FOXO3a后细胞自我更新能力的改变,以及肝癌细胞中激活IGF信号,同时过表达FOXO3a后细胞自我更新能力的改变。结果    FOXO3a在肝癌干细胞中的表达明显低于肝癌细胞(P<0.01),并且受到IGF的负向调控;构建了稳定过表达和干扰FOXO3a的细胞模型,并发现过表达FOXO3a后,细胞的克隆形成能力和成球生长能力均降低,而干扰FOXO3a后,细胞的克隆形成能力和成球生长能力均得到了提升;激活IGF信号,同时过表达FOXO3a后,细胞的克隆形成能力和成球生长能力下降。结论     FOXO3a对肝癌干细胞的自我更新能力具有负向调控作用,且可受到IGF的负调节,并介导IGF对肝癌干细胞的自我更新能力的调控。

Abstract:

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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更新日期/Last Update: 2017-08-23