HOTAIR通过STAT3/NANOG信号通路调控脑胶质瘤干细胞的增殖与分化  

HOTAIR regulates proliferation and differentiation of glioma stem cells via STAT3/NANOG signaling pathway

作　　者：石瑞 郭梦 毛维 SHI Rui;GUO Meng;MAO Wei(Clinical Laboratory,Xiunyang Central Hospital Laboratory,Xiauyann 712000,Shanxi,China;Department of Oncology,Xianyang Central Hospital,Xianyang 712000,Shanxi,China)

机构地区：[1]咸阳市中心医院检验科,陕西咸阳712000 [2]咸阳市中心医院肿瘤科,陕西咸阳712000

出　　处：《西部医学》2020年第3期355-360,共6页Medical Journal of West China

基　　金：陕西省重点研发计划(707314823001)。

摘　　要：目的探讨HOTAIR通过STAT3/NANOG信号通路对脑胶质瘤干细胞(BGSCs)的增殖与分化的调控作用。方法U251干细胞分别转入siHOTAIR(干扰组)及Control-si序列(空载组),设置空白对照组。RT-qPCR、MTT及流式细胞术检测各组细胞HOTAIR mRN A表达、细胞增殖及细胞周期分布情况;RT-qPCR及Western blot检测诱导分化后CD133、胶质酸性纤维蛋白(GFAP)、β-微血管蛋白Ⅲ(β-TublinⅢ)及髓鞘碱性蛋白(MBP)阳性细胞占比、白血病抑制因子(LIF)、STAT3、NANOG mRN A和蛋白及pSTAT3蛋白表达情况。结果与空白对照组和空载组相比,干扰组HOT AIR基因相对表达量下调,OD值降低,细胞周期G0/G1期占比降低,S、G2/M期占比升高,诱导分化后CD133、GFAP、β-TublinⅢ及MBP阳性细胞占比均较高,LIF、NANOG mRNA和蛋白相对表达量及pSTAT3/STAT3均较低(P<0.05);空白对照组与空载组上述各项指标比较差异均无统计学意义(P>0.05)。结论沉默HOTAIR基因可有效抑制BGSCs的增殖、促进分化进程,其调控机制可能与抑制LIF磷酸化激活STAT3,下调NANOG有关。Objective To investigate the regulation of proliferation and differentiation of brain glioma stem cells(BGSCs)by HOTAIR via STAT3/NANOG signaling pathway.Methods U351 stem cells were transferred into siHOTAIR and Control-si sequences,respectively.The blank control group was set up.HOTAIR gene expression,cell proliferation and cell cycle distribution were detected by RT-qPCR,MTT and flow cytometry.After induction of differentiation,the percentage of CD133,glial acid fibrin(GFAP),β-microvascular protein Ⅲ(β-Tublin Ⅲ)and myelin basic protein(MBP)positive cells,the expressions of leukemia inhibitory factor(LIF),STAT3,NANOG mRNA and protein and pSTAT3 protein were detected by RT-q PCR and Western blot.Results Compared with blank control group and no-load group,the relative expression of HOTAIR gene down-regulated,the OD value of MTT experiment decreased,the GO/G1 phase of cell cycle decreased and the S,G2/M phase increased,the percentage of CD133,GFAP,β-Tublin Ⅲ and MBP positive cells increased,while the relative expressions of LIF,NANOG mRNA and protein and pSTAT3/STAT3 decreased in the interference group(P<0.05).There were no significant differences in the above indexes between the groups(P>0.05).Conclusion Silencing HOTAIR gene can effectively inhibit the proliferation and promote the differentiation of BGSCs.Its regulatory mechanism may be related to inhibition of phosphorylation of LIF to activate STAT3 and down-regulation of NANOG mRNA and protein expression.

关 键 词：HOTAIR STAT3/NANOG信号通路 胶质瘤干细胞 增殖 分化 

分 类 号：R739.4[医药卫生—肿瘤]