Sato-Nagaoka Yuri, Suzuki Susumu, Suzuki Souma, Takahashi Shinichiro
Department of Clinical Laboratory, Tohoku Medical and Pharmaceutical University Hospital, Sendai, Japan.
Division of Laboratory Medicine, Faculty of Medicine, Tohoku Medical and Pharmaceutical University, Sendai, Japan.
PLoS One. 2024 Dec 31;19(12):e0312406. doi: 10.1371/journal.pone.0312406. eCollection 2024.
Differentiation therapy with all-trans retinoic acid (ATRA) is well established for acute promyelocytic leukemia (APL). However, the narrow application and tolerance development of ATRA remain to be improved. A number of kinase inhibitors have been reported to induce cell differentiation. In this study, we investigated several combinations of these kinase inhibitors. Recently, we revealed that the Akt inhibitor triciribine (TCN) efficiently induces differentiation of NB4 APL cells and acute myeloid leukemia (AML) M2-derived HL-60 cells through activation of the ERK/MAPK pathway. In the present study, we found that the p38 MAPK inhibitor PD169316 had profoundly enhanced the TCN effect for differentiation of NB4 and HL-60 cells. Morphologically, the combination of these two agents efficiently reduced the nuclear-to-cytoplasmic ratio and induced the expression of myelomonocytic markers (CD11b, CD11c) and some ectopic markers (erythroid glycophorin A, lymphoid CD7 and CD20), as determined by PCR and flow cytometry analyses. Western blotting analysis revealed that these agents efficiently induced phosphorylation of ERK. To clarify the molecular mechanisms involved in the TCN and PD169316-induced differentiation, we performed microarray analyses using NB4 cells. Pathway analysis using DAVID software indicated that "viral protein interaction with cytokine and cytokine receptor" and "cytokine-cytokine receptor interaction" were enriched with high significance. Real-time PCR analysis demonstrated that genes for components of these pathways, including chemokines like CCL1, CCL2, CCL3, CCL5, and CXCL8 as well as cytokines and receptors like CSF1, IL-10, IL-10RA, IL-10RB, IL-1β, and TNFSF10, were upregulated in NB4 and HL-60 cells during TCN and PD169316-induced differentiation.
全反式维甲酸（ATRA）用于急性早幼粒细胞白血病（APL）的分化治疗已得到充分确立。然而，ATRA的应用范围狭窄以及耐受性的产生仍有待改善。据报道，一些激酶抑制剂可诱导细胞分化。在本研究中，我们研究了这些激酶抑制剂的几种组合。最近，我们发现Akt抑制剂曲西立滨（TCN）通过激活ERK/MAPK途径有效诱导NB4 APL细胞和急性髓系白血病（AML）M2来源的HL-60细胞分化。在本研究中，我们发现p38 MAPK抑制剂PD169316显著增强了TCN对NB4和HL-60细胞分化的作用。从形态学上看，这两种药物的组合有效降低了核质比，并诱导了髓单核细胞标志物（CD11b、CD11c）和一些异位标志物（红细胞血型糖蛋白A、淋巴细胞CD7和CD20）的表达，这通过PCR和流式细胞术分析得以确定。蛋白质印迹分析表明，这些药物有效诱导了ERK的磷酸化。为了阐明TCN和PD169316诱导分化所涉及的分子机制，我们使用NB4细胞进行了微阵列分析。使用DAVID软件进行的通路分析表明，“病毒蛋白与细胞因子和细胞因子受体的相互作用”和“细胞因子 - 细胞因子受体相互作用”具有高度显著性富集。实时PCR分析表明，在TCN和PD169316诱导分化过程中，NB4和HL-60细胞中这些通路的组成成分基因，包括趋化因子如CCL1、CCL2、CCL3、CCL5和CXCL8以及细胞因子和受体如CSF1、IL-10、IL-10RA、IL-10RB、IL-1β和TNFSF10均上调。
