YTHDC1基因在小鼠小脑发育及人髓母细胞瘤组织中的动态表达  

Dynamic expression of YTHDC1 in mice cerebellar development and human medulloblastoma

作　　者：吴高锒 赵赋[2] 张晶[2] 李春德[2] 牛亚梅[1] 刘丕楠[2] 佟伟民[1] WU Gao-lang;ZHAO Fu;ZHANG Jing;LI Chun-de;NIU Ya-mei;LIU Pi-nan;TONG Wei-min(Department of Pathology/Molecular Pathology Center CAMS,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005;Department of Neurosurgery,Beijing Tiantan Hospital,Capital Medical University,Beijing 100050,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病理学系/中国医学科学院分子病理中心,北京100005 [2]首都医科大学附属北京天坛医院神经外科,北京100050

出　　处：《基础医学与临床》2019年第7期1001-1006,共6页Basic and Clinical Medicine

基　　金：中国医学科学院医学表观遗传学研究中心项目(2018PT31015);中国医学科学院中央级公益性科研院所基本科研业务费(2016ZX310182-2)

摘　　要：目的探讨RNA m^6A甲基化结合蛋白(YTH结构域包含蛋白1,YTHDC1)在小脑发育和髓母细胞瘤中的表达模式及其可能发挥的作用。方法分别以小鼠小脑、人髓母细胞瘤和细胞系为研究对象,在转录组测序结果的基础上,分析RNA水平表达模式;用免疫印迹和免疫组化检测YTHDC1蛋白的表达;在Daoy细胞中敲低甲基转移酶METTL3并通过m^6A-IP-qPCR方法检测YTHDC1 RNA甲基化水平及对YTHDC1蛋白丰度的影响。结果小鼠小脑发育过程中YTHDC1 RNA水平未见明显改变,蛋白表达显著下降(P<0.05)。YTHDC1在P7的小脑内颗粒神经元高表达,而在P60小脑中表达降低。与正常或癌旁组织相比,YTHDC1在肿瘤组织中RNA水平未见明显改变,蛋白水平总体偏低(P<0.05)。YTHDC1在髓母细胞瘤中RNA m^6A甲基化水平显著升高(P<0.05),而METTL3敲低引起YTHDC1 m^6A甲基化水平下降以及蛋白表达上调。结论小鼠小脑发育及人髓母细胞瘤中YTHDC1蛋白表达变化显著,这一过程受到m^6A甲基化介导的转录后调控作用,进而影响小脑发育及髓母细胞瘤的发生。Objective To study the expression pattern of RNA m^6A binding protein YTHDC1 during mouse cerebellar development and in human medulloblastoma.Methods Relative expression level of YTHDC1 RNA in the mice cerebellum,medulloblastoma and Daoy cell line were analyzed based on RNA-seq results.Western blot and immunohistochemical staining were performed to detect the expression of YTHDC1 protein.The methyltransferase METTL3 was knocked down in Daoy cells,followed by m^6A-IP-qPCR method to detect the change in YTHDC1 methylation level and the subsequent effect on the YTHDC1 protein level.Results During mice cerebellar development,the YTHDC1 expression remained constant in RNA level but showed a gradual reduction in protein level(P<0.05).Similarly,the expression of YTHDC1 protein in internal granular layer significantly decreased from post-natal day 7(P7)to P60.Comparison between medulloblastoma tissues and adjacent para-carcinoma tissues,showed no apparent change of YTHDC1 in RNA level,but a significant decrease in protein level(P<0.05).Notably,YTHDC1 RNA methylation level increased in medulloblastoma(P<0.05),and the decreased m^6A methylation level of YTHDC1 caused by METTL3 knockdown resulted in increased YTHDC1 protein expression.Conclusions In both mouse cerebellar development and human medulloblastoma,a dynamic change of YTHDC1 protein expression but not YTHDC1 RNA,is observed.The RNA m^6A modification of YTHDC1 can regulate its protein expression at post-transcriptional level,suggesting that RNA m^6A methylation of YTHDC1 may have impact on cerebellar development and medulloblastoma through regulating its protein expression.

关 键 词：RNA m^6A甲基化 YTHDC1 小脑发育 髓母细胞瘤 

分 类 号：R739.4[医药卫生—肿瘤]