Purpose: The long-term survival rate of osteosarcoma, which is the most common type of primary malignant bone tumor, has stagnated in past decades. Acacetin is a natural flavonoid compound that has antioxidative and anti-inflammatory effects and exhibits extensive therapeutic effects on various cancers. In this study, the anticancer potential of acacetin and the underlying molecular mechanisms were examined in human osteosarcoma cells (SJSA and HOS).
Materials and Methods: HOS and SJSA cell lines were exposed to different concentrations of acacetin. Cell proliferation and viability were assessed by CCK-8 and colony-formation assays. Hoechst 33258 fluorescent staining was employed to detect apoptosis. Cell apoptosis was measured by an annexin V-FITC/PI assay by flow cytometry. The alteration in the mitochondrial membrane potential was detected by a JC-1 Assay Kit. Apoptosis-related protein expression was determined by Western blotting. Intracellular reactive oxygen species (ROS) production was detected by fluorescence microscopy and flow cytometry. Subsequently, the activation of the ROS/JNK signaling pathway was investigated.
Results: Acacetin could inhibit proliferation and induce apoptosis in SJSA and HOS cells. The acacetin treatment resulted in the activation of caspase-3, − 8, and − 9 and cleaved PARP. Further studies showed that acacetin-induced apoptosis was attributed to ROS. In addition, we found that acacetin induced the activation of the downstream c-Jun N-terminal kinase (JNK) signaling pathway. Subsequently, after treatment with the ROS scavenger GSH and the JNK inhibitor SP600125, the apoptosis-inducing effect triggered by acacetin was significantly attenuated.
Conclusion: The results of the present study indicate that acacetin may induce apoptosis to inhibit cell growth by activating the ROS/JNK signaling pathway in SJSA and HOS cells, suggesting that acacetin may be a promising candidate for the management of osteosarcomas.

Keywords: acacetin, osteosarcoma, apoptosis, ROS/JNK activation


中文翻译：

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金合欢素通过调节 ROS/JNK 激活诱导人骨肉瘤细胞凋亡

目的：骨肉瘤是最常见的原发性恶性骨肿瘤类型，其长期生存率在过去几十年一直停滞不前。金合欢素是一种天然类黄酮化合物，具有抗氧化和抗炎作用，对各种癌症具有广泛的治疗作用。在这项研究中，在人骨肉瘤细胞（SJSA 和 HOS）中检查了金合欢素的抗癌潜力和潜在的分子机制。
材料和方法：HOS 和 SJSA 细胞系暴露于不同浓度的金合欢素。通过 CCK-8 和集落形成测定评估细胞增殖和活力。Hoechst 33258 荧光染色用于检测细胞凋亡。通过流式细胞术通过膜联蛋白V-FITC/PI测定法测量细胞凋亡。通过 JC-1 检测试剂盒检测线粒体膜电位的变化。通过蛋白质印迹测定凋亡相关蛋白的表达。通过荧光显微镜和流式细胞术检测细胞内活性氧 (ROS) 的产生。随后，研究了 ROS/JNK 信号通路的激活。
结果：Acacetin 可以抑制 SJSA 和 HOS 细胞的增殖并诱导细胞凋亡。金合欢素处理导致 caspase-3、-8 和 -9 的激活并切割 PARP。进一步的研究表明，金合欢素诱导的细胞凋亡归因于 ROS。此外，我们发现金合欢素诱导下游 c-Jun N-末端激酶 (JNK) 信号通路的激活。随后，在用 ROS 清除剂 GSH 和 JNK 抑制剂 SP600125 处理后，金合欢素引发的细胞凋亡诱导作用显着减弱。
结论：本研究结果表明，金合欢素可能通过激活 SJSA 和 HOS 细胞中的 ROS/JNK 信号通路诱导细胞凋亡以抑制细胞生长，表明金合欢素可能是治疗骨肉瘤的有希望的候选者。

关键词：金合欢素，骨肉瘤，细胞凋亡，ROS/JNK激活