烯醇化酶1通过影响mRNA定位调节人胚胎干细胞自我更新能力  

Enolase1 regulates the self-renewal ability of human embryonic stem cells by influencing mRNA localization

作　　者：赫佳音 杨嘉宾 陈仲扬 马艳妮[1] 余佳[1] HE Jiayin;YANG Jiabin;CHEN Zhongyang;MA Yanni;YU Jia(State Key Laboratory of Medical Molecular Biology,Department of Biochemistry and Molecular Biology,Institute of Basic Medical Science CAMS,School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所、北京协和医学院基础学院生物化学与分子生物学系、医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2023年第6期867-874,共8页Basic and Clinical Medicine

基　　金：国家自然科学基金(82122005)。

摘　　要：目的探究烯醇化酶1(ENO1)的表达对维持人胚胎干细胞(hESCs)自我更新能力的影响,并揭示其机制。方法预测与ENO1结合的RNA和蛋白质,并进行GO富集分析。在hESCs中通过shRNA抑制ENO1内源表达,集落形成实验及碱性磷酸酶染色(AP)检测hESCs的集落形成能力,荧光定量PCR检测hESCs的多能性及分化标志基因的表达变化,RNA荧光原位杂交技术检测带有polyA尾的RNA的定位变化。结果抑制ENO1的内源表达后,hESCs的集落形成能力得到了显著抑制(P<0.05),同时分化标志基因表达水平升高(P<0.001),带polyA尾的RNA的定位由分散在细胞质中变为集中在核内(P<0.001)。结论ENO1通过影响mRNA在细胞内的定位调控hESCs自我更新能力。Objective To explore the effect of enolase1(ENO1)expression on maintaining the self-renewal activity of human embryonic stem cells(hESCs)and reveals its mechanism.Methods RNAs and proteins bound to ENO1 were predicted and analyzed through Gene Oncology(GO)enrichment.Endogenous expression of ENO1 was inhibited by shRNA in hESCs.The colony forming of hESCs was examined by clonogenesis and alkaline phosphatase(AP)staining experiments.The pluripotency and differentiation marker gene expression in hESCs were detected by qPCR.RNA fluorescence in situ hybridization was used to detect the localization changes of poly(A)+RNAs.Results Inhibition of ENO1 endogenous expression significantly inhibited colony forming of hESCs(P<0.05)and the expression of differentiation marker genes was increased(P<0.001).Localization of poly(A)+RNA changed from cytoplasm to nucleus(P<0.01)with ENO1 inhibition.Conclusions ENO1 regulates the activity of hESCsself-renew by regulating mRNA localization.

关 键 词：人胚胎干细胞 烯醇化酶1 RNA结合蛋白 多能性 mRNA定位 

分 类 号：R34[医药卫生—基础医学]