成人淋巴瘤中最常见的肿瘤是弥漫性大 B 细胞淋巴瘤 (DLBCL)，其典型特征是无痛且进行性淋巴结肿大。由于 DLBCL 的高度异质性，30-40% 的患者对 R-CHOP 标准化学免疫疗法有耐药性。 DCZ0358是我们课题组以小檗碱为原料设计合成的新化合物，其抑制DLBCL生长的分子机制引起了人们的广泛关注。在本研究中，我们采用 CCK8 测定法揭示 DCZ0358 以时间和剂量依赖性方式抑制 DLBCL 细胞的增殖。此外，流式细胞术和蛋白质印迹结果显示，DCZ0358 下调 CDK4、CDK6 和 CyclinD1 的表达，从而阻断 G0/G1 期细胞周期进程。此外，DCZ0358增强线粒体膜电位去极化，促进线粒体通透性转运孔开放，增加细胞质Ca2+水平并减少细胞内三磷酸腺苷的产生，从而导致线粒体功能障碍。特别是，DCZ0358 治疗触发细胞凋亡并升高细胞内活性氧 (ROS) 水平，随后介导 JNK 通路激活。进一步的研究表明，ROS 清除剂 N-乙酰半胱氨酸 (NAC) 和 JNK 抑制剂 SP600125 预处理可以部分减弱 DCZ0358 引发的细胞凋亡和 DNA 损伤。最重要的是，当与临床常用抗 DLBCL 药物依托泊苷联合使用时，DCZ0358 在体外和体内均表现出协同抗肿瘤作用。上述结果证明了DCZ0358在DLBCL细胞中的抗肿瘤分子机制，并强调ROS/JNK/DNA损伤通路作为治疗的潜在靶点，这对开发更有效的DLBCL临床治疗方法具有重要意义。版权所有 © 2023 Elsevier B.V. All保留权利。

The most common neoplasm among adult lymphomas is diffuse large B-cell lymphoma (DLBCL), typically characterized by pain-free and progressive lymph node enlargement. Due to high heterogeneity of DLBCL, 30-40 % of patients are resistant to R-CHOP standard chemoimmunotherapy. DCZ0358 is a new compound designed and synthesized from berberine by our group and the molecular mechanism by which it inhibited DLBCL growth has attracted our widespread attention. In this study, we employed the CCK8 assay to reveal that DCZ0358 inhibited proliferation in a dependent manner of time and dosage of DLBCL cells. Moreover, flowcytometry and western blot results showed that DCZ0358 downregulated the expression of CDK4, CDK6 and CyclinD1 to block cell cycle progression in G0/G1 phase. Furthermore, DCZ0358 enhanced mitochondrial membrane potential depolarization, promoted mitochondrial permeability transport pore openness, increased cytoplastic Ca2+ levels and decreased intracellular adenosine triphosphate production, which led to mitochondrial dysfunction. In particular, DCZ0358 treatment triggered cell apoptosis and elevated intracellular reactive oxygen species (ROS) levels, which subsequently mediated JNK pathway activation. Further research indicated the pre-treatment with ROS scavenger N-acetylcysteine (NAC) and JNK inhibitor SP600125 could partially attenuate apoptosis and DNA damage triggered by DCZ0358. Most importantly, DCZ0358 exhibited synergistic anti-tumor effects when combined with etoposide, a common clinical anti-DLBCL drug, both in vitro and certainly in vivo. Above results demonstrated anti-tumor molecular mechanism of DCZ0358 in DLBCL cells and highlighted the ROS/JNK/DNA damage pathway as a potential target in therapies, which have implications for the development of more effective clinical treatments for DLBCL.Copyright © 2023 Elsevier B.V. All rights reserved.