低剂量索拉非尼联合全反式维A酸诱导野生型Fms样酪氨酸激酶3的急性髓系白血病细胞分化

doi:10.16138/j.1673-6087.2022.06.002

摘要/Abstract

摘要：

目的：研究低剂量索拉非尼与全反式维A酸（all-trans retinoic acid, ATRA）联合，对野生型Fms样酪氨酸激酶3（Fms like tyrosine kinase3, FLT3）的急性髓系白血病（acute myeloid leukemia, AML）细胞的效应及机制。方法：以野生型FLT3的AML细胞系HL-60、U937以及ATRA耐药的HL-60细胞系——HL-60Res为体外模型，以细胞表面分化抗原CD11b和形态学观察评估细胞分化；应用蛋白质印迹法研究Raf蛋白激酶、促分裂原活化的蛋白激酶（mitogenactivated protein kinase, MEK）和胞外信号调节激酶（extracellular signal-regulated kinase, ERK）的活化状态、PU.1、C/增强子结合蛋白（enhancer-binding protein, EBP）β和C/EBPε的蛋白含量。结果：低剂量（0.1~0.5 μmol/L）索拉非尼促进ATRA诱导HL-60、U937和HL-60Res 3株细胞系分化。两药联合活化Raf、MEK和ERK，并上调PU.1、C/EBPβ和C/EBPε的蛋白含量，MEK的特异性抑制剂曲美替尼可抑制两药诱导的细胞分化、MEK/ERK活化以及PU.1、C/EBPβ和C/EBPε的蛋白含量上调。结论：索拉非尼与ATRA联合，通过Raf/MEK/ERK通路上调PU.1、C/EBPβ和C/EBPε的蛋白含量，诱导野生型FLT3的AML细胞分化。

关键词: 索拉非尼, 全反式维A酸, 细胞分化, 急性髓系白血病

Abstract:

Objective To explore the effect and the mechanisms of the combination of low-dose sorafenib and all-tran retinoic acid (ATRA) in acute myeloid leukemia (AML) cells with wild type Fms like tyrosine kinase 3 (FLT3). Methods The wild type FLT3 AML cell lines HL-60, U937 and the ATRA-resistant HL-60 cell line, HL-60Res were used as in vitro models. The cell differentiation was evaluated with cell surface differentiation antigen CD11b and cellular morphology. The activation of Raf, mitogenactivated protein kinase (MEK) and extracellular signal-regulated kinase (ERK), the protein expression levels of PU.1, C/EBPβ and C/EBPε were measured by Western blotting assay. Results Low dose (0.1~0.5 μmol/L) sorafenib enhanced ATRA-induced differentiation in all three cell lines studied. The combination activated Raf, MEK and ERK, and up-regulated the levels of C/EBPβ, C/EBPε and PU.1. Addition of trametinib, a MEK specific inhibitor, suppressed the differentiation induced by sorafenib and ATRA, preventing the activation of MEK/ERK and up-regulation of the levels of C/EBPβ, C/EBPε and PU.1. Conclusions The combination of low-dose sorafenib and ATRA induced differentiation of AML cells with wild type FLT3 via RAF/MEK/ERK-mediated up-regulation of the protein levels of C/EBPβ, C/EBPε and PU.1.

Key words: Sorafenib, All-trans retinoic acid, Differentiation, Acute myeloid leukemia

中图分类号:

R733.71

卢昊, 奚会民, 李璐, 蔡循. 低剂量索拉非尼联合全反式维A酸诱导野生型Fms样酪氨酸激酶3的急性髓系白血病细胞分化[J]. 内科理论与实践, 2022, 17(06): 428-434.

LU Hao, XI Huimin, LI Lu, CAI Xun. Low-dose sorafenib combined with all-trans retinoic acid induces differentiation of acute myeloid leukemia cells with wild type Fms like tyrosine kinase 3[J]. Journal of Internal Medicine Concepts & Practice, 2022, 17(06): 428-434.

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