Singh Anjali, Pillai Lakshmi, Danes Dhanush, Umar Shweta, Balakrishnan Suresh
Department of Zoology, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, 390002, India.
Department of Zoology, Union Christian College, Aluva, Kerala, 683102, India.
Mol Biol Rep. 2025 Apr 23;52(1):415. doi: 10.1007/s11033-025-10517-z.
The tumor microenvironment plays a critical role in cancer progression, with tumor-associated macrophages regulating immune responses. These macrophages can adopt a pro-inflammatory M1 phenotype that suppresses tumor growth or an anti-inflammatory M2 phenotype that promotes progression. Reprogramming macrophages toward the M1 phenotype is a therapeutic strategy. Previous studies showed that 4-Fluorophenylacetamide-acetyl coumarin (4-FPAC), a synthetic coumarin derivative, exhibits cytostatic activity in A549 lung carcinoma cells by modulating reactive oxygen species (ROS), nitric oxide synthase, and signaling pathways, including PI3K/AKT/NF-κB. This study evaluates the impact of 4-FPAC on macrophage polarization.
We hypothesized that 4-FPAC promotes M1 macrophage polarization while suppressing M2 markers through modulation of signaling pathways, thus serving as an immunomodulatory agent.
Treatment with 4-FPAC induced M1 polarization in THP1-derived macrophages, evident from morphological elongation, elevated ROS and NO production, and increased IL-12 levels. IL-10 levels and M2 markers (CD163, STAT3, AKT1) were downregulated, while M1 markers (CD80, STAT1, AKT2) were upregulated. Gene expression and western blot analyses revealed activation of P38 and NF-κB pathways and reduced phosphorylated AKT1 levels. In silico docking showed strong interactions of 4-FPAC with regulatory proteins like P38, NF-κB, and AKT1, suggesting pathway modulation.
4-FPAC facilitates M1 macrophage polarization and inhibits M2 signaling, demonstrating its potential as an immunomodulatory agent. Coupled with its cytostatic effects on A549 cells, these findings position 4-FPAC as a promising candidate for cancer therapy. Further in vivo studies are warranted to validate its therapeutic potential and explore applications in immunotherapy and inflammation-associated diseases.
肿瘤微环境在癌症进展中起关键作用，肿瘤相关巨噬细胞调节免疫反应。这些巨噬细胞可呈现促炎性M1表型（抑制肿瘤生长）或抗炎性M2表型（促进肿瘤进展）。将巨噬细胞重编程为M1表型是一种治疗策略。先前的研究表明，4-氟苯基乙酰胺-乙酰香豆素（4-FPAC），一种合成香豆素衍生物，通过调节活性氧（ROS）、一氧化氮合酶和信号通路（包括PI3K/AKT/NF-κB）在A549肺癌细胞中表现出细胞生长抑制活性。本研究评估4-FPAC对巨噬细胞极化的影响。
我们假设4-FPAC通过调节信号通路促进M1巨噬细胞极化，同时抑制M2标志物，从而作为一种免疫调节剂。
用4-FPAC处理诱导THP1衍生的巨噬细胞发生M1极化，从形态伸长、ROS和NO产生增加以及IL-12水平升高可以明显看出。IL-10水平和M2标志物（CD163、STAT3、AKT1）下调，而M1标志物（CD80、STAT1、AKT2）上调。基因表达和蛋白质免疫印迹分析显示P38和NF-κB通路激活，磷酸化AKT1水平降低。计算机对接显示4-FPAC与P38、NF-κB和AKT1等调节蛋白有强烈相互作用，提示通路调节。
4-FPAC促进M1巨噬细胞极化并抑制M2信号传导，证明其作为免疫调节剂的潜力。再加上其对A549细胞的细胞生长抑制作用，这些发现使4-FPAC成为癌症治疗的有希望的候选药物。有必要进行进一步的体内研究以验证其治疗潜力，并探索其在免疫治疗和炎症相关疾病中的应用。
