RNA结合蛋白PCBP1通过影响mRNA核质穿梭调控hESC多能性  

RNA binding protein PCBP1 regulates hESC pluripotency through modulating mRNA export

作　　者：徐嘉悦 杨嘉宾 陈仲扬 余佳[1] 马艳妮[1] XU Jia-yue;YANG Jia-bin;CHEN Zhong-yang;YU Jia;MA Yan-ni(State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology,Institute of Basic Medicine 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》2022年第2期228-234,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金青年科学基金(31900072)。

摘　　要：目的探究RNA结合蛋白多聚胞嘧啶结合蛋白1(PCBP1)在人胚胎干细胞(hESC)多能性维持中的生物学功能,探索其调控hESC多能性的新机制。方法在hESC中敲低PCBP1蛋白进行克隆形成能力检测及碱性磷酸酶染色实验;qPCR检测多能性/分化标志基因的表达变化;同时分离hESC细胞核与细胞质进行RNA测序;对PCBP1蛋白敲低产生的RNA核质变化进行多层次分析。结果PCBP1蛋白敲低后显著抑制了人胚胎干细胞的克隆形成能力(P<0.01)并导致多能性相关基因RNA水平降低(P<0.01),细胞分化相关基因表达水平升高(P<0.01),同时显著影响了hESC中RNA整体核质分布及胚胎发育、细胞分化相关基因mRNA的核质分布。结论PCBP1通过调控多能性/分化标志基因的表达及mRNA的核质定位调控hESC多能性。Objective To investigate the biological function of RNA binding protein polycytosine-binding protein 1(PCBP1)in the maintenance of pluripotency of human embryonic stem cells(hESC)and to explore the new mechanism of regulating hESC pluripotency.Methods PCBP1 knockdown technology was used to detect the clonogenicity.The alkaline phosphatase staining profile and the expression of pluripotency/differentiation marker genes were detected by qPCR.Meanwhile,the nucleus and cytoplasm of hESC were isolated for RNA sequencing,and the nucleoplasmic changes of RNA generated by PCBP1 knockdown were analyzed by multi-level analysis.Results PCBP1 knockdown significantly inhibited clonogenesis of human embryonic stem cells(P<0.01)and reduced the RNA level of pluripotency related genes(P<0.01).The expression of differentiation related genes increased(P<0.01)that significantly affected the overall nucleoplasmic distribution of RNA in hESC and the nucleoplasmic distribution of mRNA related to embryonic development and cell differentiation.ConclusionsPCBP1 regulates hESC pluripotency by regulating the expression of pluripotency/differentiation marker genes andthe nucleoplasmic localization of mRNA.

关 键 词：人胚胎干细胞 多聚胞嘧啶结合蛋白1 多能性与自我更新 核质分离 

分 类 号：R34[医药卫生—基础医学]