β-654地中海贫血小鼠骨髓红系终末分化的阶段特异性转录组特征

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (9): 1182-1189.

β-654地中海贫血小鼠骨髓红系终末分化的阶段特异性转录组特征

史丽芳, 余东林, 刘雪会^*, 吕湘^*

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医学分子生物学国家重点实验室, 北京 100005

收稿日期:2020-06-19 修回日期:2020-06-24 出版日期:2020-09-05 发布日期:2020-09-04
通讯作者: ^*lvxiang@pumc.edu.cn;liuxuehui@ibms.pumc.edu.cn
基金资助:
国家重点研发计划干细胞及转化研究专项(2016YFA0100603);国家自然科学基金(81670108); 中国医学科学院医学与健康科技创新工程协同创新团队项目(2016-I2M-3-002)

Characterizing stage-specific transcriptome of terminal erythroid differentiation in bone marrow of β-654 thalassemia mice

SHI Li-fang, YU Dong-lin, LIU Xue-hui^*, LYU Xiang^*

State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China

Received:2020-06-19 Revised:2020-06-24 Online:2020-09-05 Published:2020-09-04
Contact: ^*lvxiang@pumc.edu.cn;liuxuehui@ibms.pumc.edu.cn

摘要/Abstract

摘要： 目的探索β-654地中海贫血小鼠骨髓红系终末分化的阶段特异性转录组特点。方法对β-654地中海贫血小鼠及同窝野生对照小鼠进行基因型鉴定及外周血涂片染色分析,而后对其骨髓细胞做流式细胞分选,收集原始红、早幼红、中幼红和晚幼红细胞用于转录组测序;测序数据过滤后进行基因差异表达分析,进一步对差异基因做GO富集分析并通过STRING数据库和Cytoscape软件对其中的上调基因进行蛋白质间相互作用网络和关键基因分析。结果 β-654地中海贫血小鼠骨髓红系细胞与同期正常对照间的差异基因随分化进程逐渐增多;GO分析显示地中海贫血早幼红-晚幼红细胞分化过程中分子伴侣介导的蛋白质折叠过程被逐渐激活,早幼红-中幼红时期氧化还原酶活性上调,中幼红-晚幼红时期泛素蛋白连接酶结合增强;蛋白质间相互作用分析发现分子伴侣中的热休克蛋白(Hsp)家族成员位于地中海贫血红系调控网络核心,且随贫血细胞向晚期分化该家族核心网络成员逐渐增多。结论 β-654地中海贫血小鼠骨髓红系细胞终末分化具有阶段特异性转录组变化,Hsp家族介导的蛋白质折叠过程随贫血细胞分化逐渐增强,早中幼红时期激活氧化还原反应,而中晚幼红时期以泛素化介导的蛋白降解为主。

关键词: 转录组, β-654地中海贫血, 热休克蛋白, 蛋白质折叠

Abstract: Objective To characterize the stage-specific transcriptome of terminal erythroid differentiation in bone marrow of β-654 thalassemia mice. Methods Genotyping and blood smear Wright-Giemsa staining were carried out to identify β-654 thalassemia mice and wild-type littermates. Bone marrow cells of β-654 thalassemia and wild-type littermates were then sorted by flow cytometry. ProE, BasoE, PolyE and OrthoE erythroblasts were collected respectively for RNA-seq; The sequencing data were filtered and subjected to differential expression analysis and subsequent GO analysis. Finally, protein-protein interaction network and hub of the up-regulated genes were depicted using STRING database and Cytoscape software. Results The quantity of differentially expressed genes between the β-654 thalassemic and stage-matched control erythroid cells increased gradually during terminal erythroid differenti- ation. GO analysis showed that chaperone-mediated protein folding process was gradually activated in thalassemic erythroid cells. Specifically, oxidoreductase activity was increased at BasoE to PolyE stages, and ubiquitin protein ligase binding was enhanced from PolyE to OrthoE stages; Protein-protein interaction analysis showed that heat shock protein (Hsp) family members were at the core of the thalassemic erythroid network and that more Hsp members join the network at later stages. Conclusions The thalassemic terminal erythroid differentiation shows stage-specific transcriptome characteristics. Protein folding mediated by Hsp family members is gradually activated during the whole process of terminal erythroid differentiation, oxidoreductase activity is selectively enhanced at BasoE and PolyE stages, while ubiquitin mediated-protein degradation is dominant during PolyE and OrthoE stages.

Key words: transcriptome, β-654 thalassemia, heat shock protein, protein folding

中图分类号:

Q811.4

史丽芳, 余东林, 刘雪会, 吕湘. β-654地中海贫血小鼠骨髓红系终末分化的阶段特异性转录组特征[J]. 基础医学与临床, 2020, 40(9): 1182-1189.

SHI Li-fang, YU Dong-lin, LIU Xue-hui, LYU Xiang. Characterizing stage-specific transcriptome of terminal erythroid differentiation in bone marrow of β-654 thalassemia mice[J]. Basic & Clinical Medicine, 2020, 40(9): 1182-1189.

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