【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CEBPA双突变急性髓系白血病的作用机制。方法:构建K562 CEBPA p30表达细胞系，Western blot测定HHT、柔红霉素、阿糖胞苷处理前后K562 CEBPA p30表达细胞系以及U937、MOLM-13细胞系中外源和内源性CEBPA蛋白表达量的变化，测定蛋白酶抑制剂和蛋白合成抑制剂对CEBPA蛋白表达量的影响。用转录组RNA-seq分析与柔红霉素处理相比，HHT处理后基因表达和通路富集的差异。结果:无论是U937和MOLM-13细胞系中的内源性CEBPA蛋白还是K562 CEBPA p30表达细胞系中的外源性CEBPA蛋白，HHT均可降低其表达量(P<0.05),而柔红霉素和阿糖胞苷无此效果。蛋白酶体抑制剂可增加CEBPA蛋白的表达量(P<0.05),而蛋白合成抑制剂可减少CEBPA蛋白的表达量(P<0.05)。HHT处理上调了K562 CEBPA p30细胞的核糖体生成相关通路，柔红霉素则不具有这种作用。结论:HHT可抑制CEBPA蛋白的合成，降低CEBPA蛋白的表达水平，这可能是HHT治疗CEBPA双突变急性髓系白血病的作用机制。更多还原

【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 CEBPA mutations. Methods: The K562 cell line expressing CEBPA p30(K562 CEBPA p30) was established. Western blot was used to determine the changes of the expression of CEBPA protein in K562 CEBPA 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 CEBPA p30 were decreased by HHT(P<0.05) while were not by DNR or Ara-C. Proteasome inhibitors can increase the expression of CEBPA protein(P<0.05) while protein synthesis inhibitors can decrease the expression of CEBPA protein(P<0.05). The ribosome biogenesis related pathways in K562 CEBPA 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 CEBPA-double-mutant AML.更多还原