Met-tRNAiMet载体蛋白在急性髓系白血病(AML)中的表达及功能

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (4): 501-507.

Met-tRNAi^Met载体蛋白在急性髓系白血病(AML)中的表达及功能

苏鹏忠, 何家驩, 于姗, 王小爽, 余佳^*

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

收稿日期:2021-01-20 修回日期:2021-02-03 出版日期:2021-04-05 发布日期:2021-04-05
通讯作者: ^*j-yu@ibms.pumc.edu.cn
基金资助:
国家自然科学基金杰出青年基金(31725013)

Expression and function of Met-tRNAi^Met carrier proteins in acute myeloid leukemia

SU Peng-zhong, HE Jia-huan, YU Shan, WANG Xiao-shuang, YU Jia^*

State Key Lab of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China

Received:2021-01-20 Revised:2021-02-03 Online:2021-04-05 Published:2021-04-05
Contact: ^*j-yu@ibms.pumc.edu.cn

摘要/Abstract

摘要： 目的探讨Met-tRNAi^Met载体蛋白eIF2A、eIF2D和MCTS1在正常造血干细胞(HSCs)和急性髓系白血病(AML)细胞中的表达,并研究上述蛋白对AML细胞增殖的影响。方法利用公共测序数据集分析Met-tRNAi^Met载体蛋白在小鼠HSCs分化不同阶段中的表达;利用单细胞转录组测序数据集分析Met-tRNAi^Met载体蛋白在人HSCs分化不同阶段中的表达;利用公共测序数据集分析Met-tRNAi^Met载体蛋白在健康人和AML患者骨髓细胞中的表达。在人急性髓细胞样白血病细胞系MOLM13中利用慢病毒传导分别抑制eIF2A或eIF2D的内源表达,检测细胞的增殖和周期变化。利用salubrinal (10 μmol/L)处理和血清饥饿的方式处理白血病细胞系使其进入应激状态,检测eIF2A和eIF2D的表达改变。结果 eIF2A和eIF2D在造血干祖细胞中的表达显著高于成熟血液细胞,并且eIF2A和eIF2D在AML患者中表达高于健康人。与之相反,MCTS1在正常造血分化过程和AML细胞中均未见表达差异。抑制内源eIF2A或eIF2D的表达后,MOLM13细胞的增殖能力受到显著抑制(P＜0.001),且细胞周期阻滞在G₂/M期(P＜0.001)。当MOLM13受到salubrinal或血清饥饿处理后,eIF2A和eIF2D的表达显著增加。结论 eIF2A和eIF2D通过促进细胞周期的正常进行调控白血病细胞的增殖。

关键词: eIF2A, eIF2D, 造血干细胞, 急性髓系白血病

Abstract: Objective To investigate the expression of Met-tRNAi^Metcarrier proteins eIF2A, eIF2D and MCTS1 in normal hematopoietic stem cells (HSCs) and acute myeloid leukemia (AML) cells, and to study the effect of carrier proteins on the functions of AML cell lines. Methods The expression of carrier proteins among multiple stages of mouse HSCs differentiation was analyzed by public datasets. The expression of carrier proteins among multiple stages of human HSCs differentiation was analyzed in single cell RNA-sequencing datasets. The expression of carrier pro- teins among AML patients and normal was analyzed in public datasets. eIF2A or eIF2D expression was inhibited by lentivirus-mediated gene transduction in AML cell line MOLM13, cell proliferation and cell cycle was analyzed.AML cell line was treated with salubrinal (10 μmol/L) and serum starvation, which make cells in stress, and the expression of eIF2A and eIF2D were detected. Results The expression of eIF2A and eIF2D in hematopoietic stem and progenitor cells (HSPCs) was higher than mature blood cells, and in AML patients was higher than normal. On the contrary, the expression of MCTS1 had no difference in normal hematopoietic differentiation process, as well as AML vs normal. eIF2A or eIF2D knock down significantly suppressed cell proliferation (P＜0.0001), as well as caused cell cycle arrest in G₂/M (P＜0.001). After MOLM13 was treated with salubrinal and serum starvation, the expression of eIF2A and eIF2D was upregulated. Conclusions eIF2A and eIF2D may regulate AML cell growth through breaking cell cycle arrest.

Key words: eIF2A, eIF2D, hematopoietic stem cells, acute myeloid leukemia

中图分类号:

苏鹏忠, 何家驩, 于姗, 王小爽, 余佳. Met-tRNAi^Met载体蛋白在急性髓系白血病(AML)中的表达及功能[J]. 基础医学与临床, 2021, 41(4): 501-507.

SU Peng-zhong, HE Jia-huan, YU Shan, WANG Xiao-shuang, YU Jia. Expression and function of Met-tRNAi^Met carrier proteins in acute myeloid leukemia[J]. Basic & Clinical Medicine, 2021, 41(4): 501-507.

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