Cai Xiao-Ya, Huang Gui-Qin, Zhou Ye-Ming, Li Deng-Ju
Department of Hematology, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan, 430030, China.
Curr Med Sci. 2025 Apr 23. doi: 10.1007/s11596-025-00042-2.
To investigate the effects of calprotectin (S100A8/A9) on the biological activity of acute myeloid leukemia (AML) cells harboring a DNA methyltransferase 3A (DNMT3A) mutation and to explore the underlying molecular mechanisms involved.
AML monoclonal cell lines harboring the DNMT3A mutation were generated via lentiviral transduction and limiting dilution. RNA sequencing was used for differential gene expression analysis, followed by bioinformatic pathway enrichment and gene correlation analyses. The biological effects of paquinimod, a selective S100A8/A9 inhibitor, on DNMT3A AML cells were assessed via Cell Counting Kit (CCK-8) proliferation assays, Annexin V/PI staining, cell cycle analysis, cell adhesion assays, and transwell migration assays.
Differential gene expression analysis revealed 442 upregulated and 535 downregulated genes in DNMT3A-mutated (DNMT3A) cells compared with those in DNMT3A wild-type (DNMT3A) cells, with the S100A8/A9 complex recurrently enriched in Reactome pathway analysis. Compared with healthy controls, patients with AML presented increased expression of S100A8 and S100A9 and increased expression of DNMT3A cells relative to DNMT3A cells, which was correlated with poor prognosis in patients with AML. There were no notable differences in proliferation among the DNMT3A, DNMT3A, and empty vector cells under normal or starvation conditions. However, paquinimod treatment notably inhibited the proliferation, migration, and adhesion of DNMT3A AML cells in a dose-dependent manner, causing G0/G1 cell cycle arrest, whereas no significant effects on apoptosis were observed. Paquinimod also downregulated key adhesion molecules, including intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), monocyte chemoattractant protein-1 (MCP-1), and matrix metalloproteinase-2 (MMP-2). Additionally, S100A8 and S100A9 expression was upregulated in a dose-dependent manner in response to cytarabine treatment.
Elevated S100A8/A9 expression contributes to the abnormal proliferation, migration, adhesion, and chemoresistance of DNMT3A AML cells. Targeting S100A8/A9 alone or in combination with other treatments represents a promising therapeutic strategy for DNMT3A AML.
研究钙卫蛋白（S100A8/A9）对携带DNA甲基转移酶3A（DNMT3A）突变的急性髓系白血病（AML）细胞生物学活性的影响，并探讨其潜在的分子机制。
通过慢病毒转导和有限稀释法建立携带DNMT3A突变的AML单克隆细胞系。采用RNA测序进行差异基因表达分析，随后进行生物信息通路富集和基因相关性分析。通过细胞计数试剂盒（CCK-8）增殖试验、膜联蛋白V/碘化丙啶染色、细胞周期分析、细胞黏附试验和Transwell迁移试验，评估选择性S100A8/A9抑制剂帕喹莫德对DNMT3A AML细胞的生物学作用。
差异基因表达分析显示，与DNMT3A野生型（DNMT3A）细胞相比，DNMT3A突变型（DNMT3A）细胞中有442个基因上调，535个基因下调，在Reactome通路分析中，S100A8/A9复合物反复富集。与健康对照相比，AML患者的S100A8和S100A9表达增加，且与DNMT3A细胞相比，DNMT3A细胞的表达增加，这与AML患者的不良预后相关。在正常或饥饿条件下，DNMT3A、DNMT3A和空载体细胞之间的增殖没有显著差异。然而，帕喹莫德治疗以剂量依赖的方式显著抑制DNMT3A AML细胞的增殖、迁移和黏附，导致G0/G1期细胞周期阻滞，而未观察到对细胞凋亡的显著影响。帕喹莫德还下调了关键黏附分子，包括细胞间黏附分子1（ICAM-1）、血管细胞黏附分子1（VCAM-1）、单核细胞趋化蛋白-1（MCP-1）和基质金属蛋白酶-2（MMP-2）。此外，在阿糖胞苷治疗后，S100A8和S100A9的表达呈剂量依赖性上调。
S100A8/A9表达升高促进了DNMT3A AML细胞的异常增殖、迁移、黏附和化疗耐药。单独靶向S100A8/A9或与其他治疗联合使用是DNMT3A AML一种有前景的治疗策略。
