Ahmadi Abdolreza, Hosseini Fatemehsadat, Iranshahy Milad, Rassouli Fatemeh B
Novel Diagnostics and Therapeutics Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
Sci Rep. 2025 Jul 17;15(1):25941. doi: 10.1038/s41598-025-11804-2.
Osteosarcoma is an aggressive malignancy marked by a high incidence of local recurrence and distant metastasis, leading to poor outcomes in advanced stages. While current therapies offer long-term survival primarily for patients with localized disease, effective treatments for metastatic cases remain elusive. Addressing this critical gap, the present study explores, for the first time, the anti-metastatic potential of urolithin A (UA), a naturally derived polyphenol, against osteosarcoma cells. Interactome mapping, gene enrichment profiling, target gene expression assessment, and molecular docking coupled with dynamics simulations were performed to elucidate the mechanistic basis of UA action. For experimental studies, UA was synthesized and its effects on osteosarcoma cell viability, apoptosis, migration, adhesion, invasion, and MMP activity were evaluated using alamarBlue assay, flow cytometry, scratch assay, fibronectin-based adhesion assay, Boyden chamber assay, and gelatin zymography, respectively. Results identified AKT1, EGFR, and MMP9 as potential targets of UA associated with osteosarcoma progression. Further analyses revealed critical interactions among these hub targets, with significant upregulation of AKT1 observed in osteosarcoma tissue samples. Molecular docking and dynamics simulations demonstrated strong and stable binding of UA to the kinase domain of AKT1 and the active site of EGFR. Experimental validation showed that treatment with UA significantly inhibited the migration and invasion of osteosarcoma cells, while notably enhancing cell adhesion. This anti-metastatic effect was closely linked to a marked reduction in enzymatic activity of MMP2 and MMP9, key mediators of metastatic dissemination. These findings position UA as a promising therapeutic candidate for targeting osteosarcoma metastasis.
骨肉瘤是一种侵袭性恶性肿瘤，其特征是局部复发和远处转移的发生率很高，导致晚期预后不良。虽然目前的治疗方法主要为局限性疾病患者提供长期生存，但转移性病例的有效治疗方法仍然难以捉摸。为了填补这一关键空白，本研究首次探索了天然衍生的多酚乌罗托品A（UA）对骨肉瘤细胞的抗转移潜力。进行了相互作用组图谱绘制、基因富集分析、靶基因表达评估以及结合动力学模拟的分子对接，以阐明UA作用的机制基础。在实验研究中，合成了UA，并分别使用alamarBlue测定法、流式细胞术、划痕试验、基于纤连蛋白的粘附试验、Boyden小室试验和明胶酶谱法评估了其对骨肉瘤细胞活力、凋亡、迁移、粘附、侵袭和MMP活性的影响。结果确定AKT1、EGFR和MMP9是与骨肉瘤进展相关的UA潜在靶点。进一步分析揭示了这些核心靶点之间的关键相互作用，在骨肉瘤组织样本中观察到AKT1显著上调。分子对接和动力学模拟表明UA与AKT1的激酶结构域和EGFR的活性位点有强烈且稳定的结合。实验验证表明，用UA处理可显著抑制骨肉瘤细胞的迁移和侵袭，同时显著增强细胞粘附。这种抗转移作用与转移扩散的关键介质MMP2和MMP9的酶活性显著降低密切相关。这些发现使UA成为靶向骨肉瘤转移的有前途的治疗候选物。
