Goudarzi Sajad, Nasiri Sarvi Zahra, Ayatollahi Hossein, Ghodsi Razieh, Noroozi Zeinab, Keramati Mohammad Reza, Rassouli Fatemeh B
Novel Diagnostics and Therapeutics Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Cancer Molecular Pathology Research Center, Department of Hematology and Blood Bank, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Med Oncol. 2025 Jun 17;42(7):270. doi: 10.1007/s12032-025-02793-3.
Adult T-cell leukemia/lymphoma (ATLL) has low overall survival, underscoring the need for the development of novel approaches. Present study aimed to investigate anti-proliferative effects of entinostat and its newly synthesized analogs on ATLL cells. Computational analyses were conducted to identify the potential molecular targets of entinostat, and construct a protein-protein interaction network. Then, enrichment analyses were performed, and the expression of CDK4 was assessed in MT-2 cells. Molecular docking and dynamics simulations were carried out to predict the interactions of entinostat and its novel analogs with target proteins. For in vitro studies, at first quinoline-based benzamide derivatives were synthesized. Then, MT-2 and normal cells were treated and their proliferation was evaluated by alamarBlue assay. Finally, flow cytometry was performed, and the expression of candidate genes was assessed by real-time PCR. Exploring potential targets of entinostat and pathogenic targets of ATLL revealed 51 overlapping molecules including CDK4. Volcano plot revealed over expression of CDK4 in MT-2 cells. Favorable and stable binding of entinostat and its analogs with the activation loop of CDK4 and the CDK-binding site of cyclin D1 was confirmed. Experimental studies revealed anti-proliferative effects of entinostat and analogs on MT-2 cells, confirmed by flow cytometry analysis and alterations in the expression of BAX, CCND1, and BCL-2. Present findings pave the way for the development of new drugs against ATLL, and provide evidence that justifies further preclinical evaluations of entinostat and its novel analogs.
成人T细胞白血病/淋巴瘤(ATLL)的总体生存率较低,这突出了开发新方法的必要性。本研究旨在探讨恩替诺特及其新合成类似物对ATLL细胞的抗增殖作用。进行了计算分析以确定恩替诺特的潜在分子靶点,并构建蛋白质-蛋白质相互作用网络。然后,进行富集分析,并在MT-2细胞中评估CDK4的表达。进行了分子对接和动力学模拟,以预测恩替诺特及其新型类似物与靶蛋白的相互作用。对于体外研究,首先合成了喹啉基苯甲酰胺衍生物。然后,处理MT-2细胞和正常细胞,并通过alamarBlue测定法评估它们的增殖。最后,进行流式细胞术,并通过实时PCR评估候选基因的表达。探索恩替诺特的潜在靶点和ATLL的致病靶点发现了51个重叠分子,包括CDK4。火山图显示MT-2细胞中CDK4的过表达。证实了恩替诺特及其类似物与CDK4的激活环和细胞周期蛋白D1的CDK结合位点具有良好且稳定的结合。实验研究揭示了恩替诺特及其类似物对MT-2细胞的抗增殖作用,流式细胞术分析以及BAX、CCND1和BCL-2表达的改变证实了这一点。目前的研究结果为开发抗ATLL的新药铺平了道路,并提供了证据证明对恩替诺特及其新型类似物进行进一步临床前评估是合理的。