摘要
目的探究粒细胞特异表达的HOXA转录本反义RNA1(HOTAIRM1)在胶质瘤干细胞中发挥的功能及分子机制。方法Real-timePCR检测HOTAIRM1在胶质瘤干细胞中的表达谱;肿瘤球形成实验和有限稀释实验检测过表达或敲低HOTAIRM1后胶质瘤干细胞自我更新能力;real-timePCR和Westernblot检测干性标志物的表达;双荧光素酶报告基因实验检测OCT4启动子活性。全反式维甲酸(ATRA)处理NB4急性早幼粒细胞白血病细胞和胶质瘤干细胞GSC2后检测HOTAIRM1表达。结果与对照组相比,过表达HOTAIRM1的胶质瘤干细胞自我更新能力呈显著上升(P<0.05);干性标志物NESTIN、OCT4及SOX2表达增加(P<0.05)。敲低HOTAIRM1则结果相反。过表达HOTAIRM1可促进OCT4启动子转录活性(P<0.05)。ATRA处理GSC2后HOTAIRM1表达下调(P<0.01)。结论HOTAIRM1通过调节OCT4表达维持胶质瘤干细胞的自我更新。
Objective To explore the function and molecular mechanism of HOXA transcript antisense RNA(myeloid-specific 1, HOTAIRM1) in glioma stem cells. Methods Real-time PCR was used to detecte the expression profile of HOTAIRM1 in glioma stem cells;tumor spheroid formation assay and limiting dilution assay were used to detect the self-renewal of tumor stem cells after HOTAIRM1 overexpression or knockdown;real time PCR and Western blot were used to detect the expression of stemness markers;dual-luciferase reporter gene assay was used to find the effect of HOTAIRM1 on the activity of OCT4 promoter;all-trans retinoic acid(ATRA) was used to treat NB4 acute promyelocytic leukemia cells and glioma stem cells GSC2. Results Compared with the control group, the self-renewal ability of glioma stem cells overexpressing HOTAIRM1 was significantly enhanced(P<0.05);Western blot showed significant up-regulation of the stemness markers(P<0.05). The results of knoc-king down HOTAIRM1 were opposite. Overexpression of HOTAIRM1 promoted the transcriptional activity of the OCT4 promoter. The expression of HOTAIRM1 was down-regulated after ATRA treatment of GSC2(P<0.01).Conclusions HOTAIRM1 maintains self-renewal of glioma stem cells by modulating OCT4 expression.
作者
王珊珊
彭小忠
韩为
WANG Shan-shan;PENG Xiao-zhong;HAN Wei(State Key Laboratory of Medical Molecular Biology,Department of Molecular Biology and Biochemistry,Medical Primate Research Center,Neuroscience Center,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005;Institute of Medical Biology,Chinese Academy of Medical Sciences,Kunming 650118,China)
出处
《基础医学与临床》
CSCD
2019年第6期792-797,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(31671316)
