School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 211816, People's Republic of China.
School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 211816, People's Republic of China.
Phytomedicine. 2024 Sep;132:155716. doi: 10.1016/j.phymed.2024.155716. Epub 2024 May 9.
Osteosarcoma (OS), the most prevalent primary bone malignancy, exhibits rapid growth and a high tendency for lung metastasis, posing significant treatment challenges. Ziyuglycoside II (ZGS II), a main active compound derived from Sanguisorba officinalis l., has shown potential in cancer treatment. However, the effects of ZGS II and its potential mechanism in OS remain elusive.
This study aims to explore the anti-metastatic potential of ZGS II in OS, offering a novel therapeutic strategy for improved patient outcomes.
Cell viability and proliferation was detected by cell counting kit-8 (CCK-8) and clone formation assay, respectively. Transwell and wound-healing assay were applied to evaluate the potential metastatic abilities of OS cells in vitro. More critically, the chromobox protein homolog 4 (CBX4) and Wnt/β-catenin signaling pathway was investigated utilizing Western blotting, immunohistochemistry, shRNA knockdown and immunofluorescence. An orthotopic metastasis mouse model was utilized to evaluate the efficacy of ZGS II in suppressing OS metastasis in vivo, with molecular docking studies conducted to elucidate the interaction between ZGS II and the CBX4 protein.
Our study demonstrated the potent inhibitory effects of ZGS II on OS cell proliferation and induced apoptosis in vitro, as evidenced by decreased cell viability, enhanced caspase-3 activation, and mitochondrial dysfunction. Furthermore, using an orthotopic metastasis mouse model, we illustrated that ZGS II effectively suppressed tumor growth and lung metastasis in vivo. Notably, our investigation revealed that the antitumor action of ZGS II is dependent on the reduction of CBX4 levels, leading to the attenuation of the Wnt/β-catenin signaling pathway activation. Molecular docking analyses supported this pathway's suppression, showing that ZGS II has the capability to directly bind and disrupt CBX4 function. To further confirm this mechanism, we utilized shRNA to silence CBX4 in OS cells, which significantly enhanced the inhibitory impact of ZGS II on cell migration.
Our study findings reveal that ZGS II efficiently suppresses both metastasis and tumor growth in OS by a novel mechanism that entails the inhibition of the CBX4-regulated Wnt/β-catenin pathway. These outcomes highlight the promising potential of ZGS II as a therapeutic agent for managing metastatic OS, thus justifying the need for additional clinical investigations.
骨肉瘤（OS）是最常见的原发性骨恶性肿瘤，具有快速生长和高肺转移倾向，治疗极具挑战性。梓醇（ZGS II）是从苦荬菜中提取的主要活性化合物，在癌症治疗中显示出潜力。然而，ZGS II 的作用及其在 OS 中的潜在机制仍不清楚。
本研究旨在探讨 ZGS II 在 OS 中的抗转移潜力，为改善患者预后提供新的治疗策略。
通过细胞计数试剂盒-8（CCK-8）和克隆形成实验分别检测细胞活力和增殖。Transwell 和划痕愈合实验用于评估 OS 细胞体外转移的潜在能力。更重要的是，利用 Western blot、免疫组化、shRNA 敲低和免疫荧光实验研究了染色质盒蛋白同源物 4（CBX4）和 Wnt/β-catenin 信号通路。利用原位转移小鼠模型评估 ZGS II 体内抑制 OS 转移的疗效，并进行分子对接研究以阐明 ZGS II 与 CBX4 蛋白的相互作用。
本研究表明 ZGS II 可有效抑制 OS 细胞增殖并诱导细胞凋亡，表现为细胞活力降低、半胱天冬酶-3 激活增强和线粒体功能障碍。此外，通过原位转移小鼠模型，我们证明 ZGS II 可有效抑制肿瘤生长和体内肺转移。值得注意的是，我们的研究发现 ZGS II 的抗肿瘤作用依赖于 CBX4 水平的降低，从而减弱 Wnt/β-catenin 信号通路的激活。分子对接分析支持该通路的抑制作用，表明 ZGS II 具有直接结合并破坏 CBX4 功能的能力。为了进一步证实这一机制，我们利用 shRNA 沉默 OS 细胞中的 CBX4，这显著增强了 ZGS II 对细胞迁移的抑制作用。
本研究结果表明，ZGS II 通过一种新的机制有效抑制 OS 中的转移和肿瘤生长，该机制涉及抑制 CBX4 调节的 Wnt/β-catenin 通路。这些结果突显了 ZGS II 作为治疗转移性 OS 的治疗剂的有前途的潜力，因此需要进一步的临床研究。
