Chinese People's Liberation Army Medical School, Beijing, 100853, China.
School of Basic Medicine, Shanxi Medical University, Taiyuan, 030000, China.
Sci Rep. 2024 Jul 4;14(1):15406. doi: 10.1038/s41598-024-66367-5.
Patients with multiple myeloma (MM) experience relapse and drug resistance; therefore, novel treatments are essential. Clotrimazole (CTZ) is a wide-spectrum antifungal drug with antitumor activity. However, CTZ's effects on MM are unclear. We investigated CTZ's effect on MM cell proliferation and apoptosis induction mechanisms. CTZ's effects on MM.1S, NCI- H929, KMS-11, and U266 cell growth were investigated using Cell Counting Kit-8 (CCK-8) assay. The apoptotic cell percentage was quantified with annexin V-fluorescein isothiocyanate/7-amino actinomycin D staining. Mitochondrial membrane potential (MMP) and cell cycle progression were evaluated. Reactive oxygen species (ROS) levels were measured via fluorescence microscopy. Expression of apoptosis-related and nuclear factor (NF)-κB signaling proteins was analyzed using western blotting. The CCK-8 assay indicated that CTZ inhibited cell proliferation based on both dose and exposure time. Flow cytometry revealed that CTZ decreased apoptosis and MMP and induced G0/G1 arrest. Immunofluorescence demonstrated that CTZ dose-dependently elevated in both total and mitochondrial ROS production. Western blotting showed that CTZ enhanced Bax and cleaved poly ADP-ribose polymerase and caspase-3 while decreasing Bcl-2, p-p65, and p-IκBα. Therefore, CTZ inhibits MM cell proliferation by promoting ROS-mediated mitochondrial apoptosis, inducing G0/G1 arrest, inhibiting the NF-κB pathway, and has the potential for treating MM.
患有多发性骨髓瘤 (MM) 的患者会经历复发和耐药；因此，新型治疗方法至关重要。克霉唑 (CTZ) 是一种具有抗肿瘤活性的广谱抗真菌药物。然而，CTZ 对 MM 的作用尚不清楚。我们研究了 CTZ 对 MM 细胞增殖和诱导凋亡的机制。用细胞计数试剂盒-8 (CCK-8) 测定 CTZ 对 MM.1S、NCI-H929、KMS-11 和 U266 细胞生长的影响。用膜联蛋白 V-荧光素异硫氰酸酯/7-氨基放线菌素 D 染色定量凋亡细胞百分比。评估线粒体膜电位 (MMP) 和细胞周期进程。通过荧光显微镜测量活性氧 (ROS) 水平。用 Western blot 分析凋亡相关和核因子 (NF)-κB 信号蛋白的表达。CCK-8 测定表明 CTZ 基于剂量和暴露时间抑制细胞增殖。流式细胞术显示 CTZ 降低凋亡和 MMP 并诱导 G0/G1 期阻滞。免疫荧光显示 CTZ 剂量依赖性地增加总 ROS 和线粒体 ROS 的产生。Western blot 显示 CTZ 增强 Bax 和裂解多聚 ADP-核糖聚合酶和半胱天冬酶-3，同时降低 Bcl-2、p-p65 和 p-IκBα。因此，CTZ 通过促进 ROS 介导的线粒体凋亡、诱导 G0/G1 期阻滞、抑制 NF-κB 通路来抑制 MM 细胞增殖，具有治疗 MM 的潜力。
