Department of Orthopedics, Ningbo No. 9 Hospital, Ningbo, Zhejiang, China.
Department of Orthopedics, Ningbo No. 2 Hospital, Ningbo, Zhejiang, China.
Anat Rec (Hoboken). 2019 Nov;302(11):1941-1949. doi: 10.1002/ar.24200. Epub 2019 Jul 16.
Osteosarcoma (OS) is one of the most common malignant bone tumors among children and young adults. Furowanin A (Fur A), one of the active ingredients of Millettia pachycarpa Benth, has been found to exert pro-apoptotic activity in human leukemia cells. This study is designed to evaluate the efficacy of Fur A against OS. The effect of Fur A on cell viability was assessed by Cell Counting Kit-8 (CCK-8) assay. Western blotting and quantitative real-time PCR (qRT-PCR) were performed to determine the protein and mRNA level of sphingosine kinase 1 (SphK1), respectively. To validate the role of SphK1 in the pro-apoptotic activity of Fur A, overexpressing SphK1 vector and siRNA targeting SphK1 were utilized to transfect OS cells. Moreover, an OS xenograft murine model was used to analyze the therapeutic efficacy of Fur A in vivo. Fur A treatment led to a dose-dependent decrease in the number of viable cells. It also exhibited antiproliferative activity and significantly promoted apoptotic cell death in OS cell lines. Our results showed that the anticancer activity of Fur A was associated with downregulation of SphK1 and inactivation of its downstream signaling. The mediatory role of SphK1 was validated when the pro-apoptotic activity of Fur A was significantly blocked by SphK1 overexpression, while SphK1 knockdown sensitized the OS cells to Fur A. We concluded that Fur A can exhibit anti-growth and pro-apoptotic activities in vitro and in vivo in OS by downregulating SphK1. Our study highlights the possibility of utilizing Fur A as a chemotherapeutic agent in the treatment of OS. Anat Rec, 302:1941-1949, 2019. © 2019 American Association for Anatomy.
骨肉瘤（OS）是儿童和青少年中最常见的恶性骨肿瘤之一。从密花豆中分离得到的活性成分呋喃阿霉素 A（Fur A）已被发现对人白血病细胞具有促凋亡活性。本研究旨在评估 Fur A 对 OS 的疗效。通过 Cell Counting Kit-8（CCK-8）检测试剂盒评估 Fur A 对细胞活力的影响。Western blot 和定量实时 PCR（qRT-PCR）分别用于测定 Sphingosine kinase 1（SphK1）的蛋白和 mRNA 水平。为了验证 SphK1 在 Fur A 促凋亡活性中的作用，利用 SphK1 过表达载体和 SphK1 靶向 siRNA 转染 OS 细胞。此外，还利用 OS 异种移植小鼠模型分析 Fur A 的体内治疗效果。Fur A 处理导致活细胞数量呈剂量依赖性减少。它还表现出抗增殖活性，并显著促进 OS 细胞系中的凋亡细胞死亡。我们的结果表明，Fur A 的抗癌活性与 SphK1 的下调及其下游信号失活有关。当 SphK1 过表达显著阻断 Fur A 的促凋亡活性时，SphK1 的中介作用得到验证，而 SphK1 敲低则使 OS 细胞对 Fur A 敏感。我们得出结论，Fur A 可以通过下调 SphK1 在体外和体内表现出抗生长和促凋亡活性。我们的研究强调了利用 Fur A 作为骨肉瘤治疗中的化疗药物的可能性。解剖学记录，302：1941-1949，2019。©2019 年美国解剖学会。
