miR-34a通过Notch1信号通路对肺癌干细胞的抑制  

Inhibitory effect of mi R-34a on lung cancer stem cells via Notch1 signaling pathway

作　　者：韩纪昌[1] 张祎捷[1] 李红兵[1] 杨存葆[1] 马超楠[1] 戚冠斌[1] 

机构地区：[1]河南大学淮河医院呼吸内科,河南省开封市475000

出　　处：《中国组织工程研究》2016年第23期3349-3356,共8页Chinese Journal of Tissue Engineering Research

基　　金：河南省卫生厅中青年科技创新人才工程(4189号)~~

摘　　要：背景:已有报道证实mi R-34a对肺癌干细胞有一定的抑制作用,但是其作用机制目前还不清楚。目的:探索mi R-34a对肺癌干细胞的抑制作用及机制。方法:应用免疫磁珠分选细胞技术从肺腺癌A549细胞系中分离出CD133+肺癌干细胞;脂质体转染技术构建miR-34a过表达的肺癌干细胞;双荧光素酶报告基因分析miR-34a与Notch1的靶向关系;基因敲除Notch1并检测其对肺癌干细胞的影响。结果与结论:(1)经过免疫磁珠分选和流式细胞术检测得到高比率的CD133+肺癌干细胞;(2)qR T-PCR检测发现miR-34a在CD133+肺癌干细胞中的表达明显低于CD133-肺癌细胞;(3)成功构建miR-34a过表达的肺癌干细胞,发现miR-34a能够抑制肺癌干细胞的增殖,诱导细胞凋亡;(4)双荧光素酶报告基因分析证明Notch1与miR-34a具有靶向关系;(5)基因敲除Notch1显著抑制肺癌干细胞的增殖,诱导细胞凋亡;(6)结果提示,miR-34a可能通过抑制Notch1信号通路来抑制肺癌干细胞的生长。BACKGROUND： It has been proved that mi R-34 a plays an inhibitory role in the growth of lung cancer stem cells, but the underlying mechanism remains unclear. OBJECTIVE： To explore the inhibitory effect of mi R-34 a on lung cancer stem cells and the underlying mechanism. METHODS： The CD133＋ lung cancer stem cells were separated from lung cancer A549 cell lines using magnetic activated cell sorting method. And miR-34a-overexpressing CD133＋ lung cancer stem cells were established by liposome transfection technology. Besides, the targeted relationship between mi R-34 a and Notch1 was analyzed by the dual-luciferase reporter. Afterwards, Notch1 silencing was performed by gene knockout, and its effect on lung cancer stem cells was determined. RESULTS AND CONCLUSION： After sorted and detected by immunomagetic selection and flow cytometry assay respectively, a high rate of CD133＋ lung cancer stem cell was obtained. And qR T-PCR detected that the expression level of mi R-34 a in CD133＋ lung cancer stem cells was significantly lower than that in CD133- lung cancer stem cells. Moreover, mi R-34a-overexpressing CD133＋ lung cancer stem cells were successfully constructed and mi R-34 a significantly inhibited proliferation and induced apoptosis of lung cancer stem cells. Dual-luciferase reporter assay indicated that Notch1 m RNA was a target of miR-34 a. In addition, Notch1 silencing obviously inhibited proliferation and induced apoptosis of lung cancer stem cells. These findings suggest that mi R-34 a can inhibite lung cancer stem cells via the Notch1 signaling pathway.

关 键 词：肺肿瘤 肿瘤干细胞 微RNAS 受体 Notch1 组织工程 干细胞 肺癌干细胞 mi RNA MIR-34A Notch1信号通路 NOTCH1 肺腺癌A549细胞 CD133+肺癌干细胞 增殖 凋亡 

分 类 号：R394.2[医药卫生—医学遗传学]