高三尖杉酯碱对CEBPA蛋白表达的影响及机制

[Effects and Mechanisms of Homoharringtonine on Expression of CEBPA Protein].

作者信息

Li Shu-Xin, Chen Jia-Yuan, Hui Yan, Rao Qing, Wang Min, Wang Jian-Xiang, Wei Hui

机构信息

State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China.

Tianjin Institutes of Health Science， Tianjin 301600, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023;31(5):1257-1262. doi: 10.19746/j.cnki.issn.1009-2137.2023.05.001.

DOI:10.19746/j.cnki.issn.1009-2137.2023.05.001

PMID:37846669

Abstract

OBJECTIVE

To investigate the effect of homoharringtonine (HHT) on CEBPA protein and explore the mechanism of HHT in the treatment of acute myeloid leukemia (AML) with double mutations.

METHODS

The K562 cell line expressing p30 (K562 p30) was established. Western blot was used to determine the changes of the expression of CEBPA protein in K562 p30, U937 and MOLM-13 cell lines before and after treatments with HHT, daunorubicin (DNR) or cytarabine (Ara-C). The effects of protease inhibitors and protein synthesis inhibitors on the expression of CEBPA protein were also determined. RNA-seq was used to analyze the difference of gene expressions and pathway enrichments between HHT group and DNR group.

RESULTS

Both the endogenous CEBPA protein in U937 and MOLM-13 cell lines and the exogenous CEBPA protein in K562 p30 were decreased by HHT (<0.05) while were not by DNR or Ara-C. Proteasome inhibitors can increase the expression of CEBPA protein (<0.05) while protein synthesis inhibitors can decrease the expression of CEBPA protein (<0.05). The ribosome biogenesis related pathways in K562 p30 were upregulated in HHT group while were not in DNR group.

CONCLUSION

HHT can inhibit the synthesis of CEBPA and reduce the expression of CEBPA protein and this may be the mechanism of HHT in the treatment of -double-mutant AML.

摘要

目的

研究高三尖杉酯碱（HHT）对CEBPA蛋白的影响，探讨HHT治疗双突变急性髓系白血病（AML）的机制。

方法

建立表达p30的K562细胞系（K562 p30）。采用蛋白质免疫印迹法检测HHT、柔红霉素（DNR）或阿糖胞苷（Ara-C）处理前后K562 p30、U937和MOLM-13细胞系中CEBPA蛋白表达的变化。还检测了蛋白酶抑制剂和蛋白质合成抑制剂对CEBPA蛋白表达的影响。采用RNA测序分析HHT组和DNR组之间基因表达差异和通路富集情况。

结果

HHT可降低U937和MOLM-13细胞系中的内源性CEBPA蛋白以及K562 p30中的外源性CEBPA蛋白（<0.05），而DNR或Ara-C则无此作用。蛋白酶体抑制剂可增加CEBPA蛋白表达（<0.05），而蛋白质合成抑制剂可降低CEBPA蛋白表达（<0.05）。HHT组K562 p30中核糖体生物合成相关通路上调，而DNR组则无此现象。