转录因子Ikzf1促进小鼠胎肝来源红系细胞终末分化

doi:10.16352/j.issn.1001-6325.2023.06.0916

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (6): 916-922.doi: 10.16352/j.issn.1001-6325.2023.06.0916

转录因子Ikzf1促进小鼠胎肝来源红系细胞终末分化

王佳鑫, 余东林, 杨希, 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病理生理学系医学分子生物学国家重点实验室,北京 100005

收稿日期:2023-03-14 修回日期:2023-04-19 出版日期:2023-06-05 发布日期:2023-05-31
通讯作者: ^*lvxiang@pumc.edu.cn; liuxuehui@ibms.pumc.edu.cn
基金资助:
国家自然科学基金(82170120)

Ikzf1 promotes terminal differentiation of mouse fetal liver derived erythroid cells

WANG Jiaxin, YU Donglin, YANG Xi, LIU Xuehui^*, LYU Xiang^*

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

Received:2023-03-14 Revised:2023-04-19 Online:2023-06-05 Published:2023-05-31
Contact: ^*lvxiang@pumc.edu.cn; liuxuehui@ibms.pumc.edu.cn

摘要/Abstract

摘要： 目的通过分析单细胞转录组数据,挖掘红系终末分化过程中潜在的转录因子Ikzf1,并探究其在终末红系分化中的功能。方法利用SCENIC方法对人和小鼠红系细胞的单细胞转录组数据进行转录因子预测,从bulk转录组数据获取所预测因子Ikzf1在红系终末分化过程中的表达情况并在小鼠胎肝红系分化体系中进行RT-qPCR验证。在小鼠胎肝来源的红系细胞中利用短发夹RNA (shRNA)干扰技术敲低Ikzf1,流式分选GFP阳性的成功转染细胞,RT-qPCR验证其敲低效率,流式细胞术分析Ikzf1敲低后对红系分化和脱核的影响。结果 SCENIC预测Ikzf1是人与小鼠红系终末分化共同的转录因子,转录组及RT-qPCR分析显示其在终末分化较早期的原红和早幼红细胞阶段表达量较高。将小鼠胎肝来源红系细胞中Ikzf1基因的表达成功敲低,流式细胞术检测发现Ikzf1敲低导致红细胞分化阻滞在早幼红细胞时期,细胞脱核率显著降低 (P＜0.05)。结论转录因子Ikzf1在红系终末分化早期高表达并参与促进红系终末分化和脱核。

关键词: Ikzf1, 红系终末分化, 红系脱核

Abstract: Objective To explore potential function of IKAROS family zinc finger 1 (Ikzf1) in terminal erythroid differentiation. Methods Single-cell regulatory network inference and clustering (SCENIC) was used to predict transcription factors of terminal erythroid differentiation based on single cell transcriptomes of human and mouse bone marrow erythroid cells. Expression of the predicted Ikzf1 during terminal erythroid differentiation was acquired from public RNA-seq data and validated by RT-qPCR. Short hairpin RNA (shRNA) interference was performed to knock down Ikzf1 in mouse erythroid cells from fetal liver with a GFP cassette to mark successfully transfected cells. The knockdown efficiency in GFP⁺ cells was verified by RT-qPCR. The effect of Ikzf1 knockdown on erythroid differentiation and enucleation was analyzed by flow cytometry. Results SCENIC predicted that the transcription factor Ikzf1 regulated both human and mouse erythroid differentiation. RNA-seq data and RT-qPCR showed highly expressed Ikzf1 at proerythroblast and basophilic erythroblast stages of terminal erythroid differentiation. Ikzf1 expression was indeed down-regulated by shRNA in erythroid cells from mouse fetal liver. Flow cytometry analysis showed that knockdown of Ikzf1 affected terminal differentiation and significantly reduced the ratio of erythroid enucleation (P＜ 0.05). Conclusions Transcription factor Ikzf1 is highly expressed in early stage erythroblasts and involved in terminal erythroid differentiation as well as enucleation.

Key words: Ikzf1, terminal erythroid differentiation, enucleation

中图分类号:

王佳鑫, 余东林, 杨希, 刘雪会, 吕湘. 转录因子Ikzf1促进小鼠胎肝来源红系细胞终末分化[J]. 基础医学与临床, 2023, 43(6): 916-922.

WANG Jiaxin, YU Donglin, YANG Xi, LIU Xuehui, LYU Xiang. Ikzf1 promotes terminal differentiation of mouse fetal liver derived erythroid cells[J]. Basic & Clinical Medicine, 2023, 43(6): 916-922.

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转录因子Ikzf1促进小鼠胎肝来源红系细胞终末分化

Ikzf1 promotes terminal differentiation of mouse fetal liver derived erythroid cells

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