Huang Huaming, Nie Chao, Qin Xiaokang, Zhou Jie, Zhang Lei
Department of Research Office, Jiangsu Health Vocational College, Nanjing, Jiangsu 211800, P.R. China.
Department of Orthopedics, Xishan People's Hospital of Wuxi, Wuxi, Jiangsu 214015, P.R. China.
Oncol Lett. 2019 Oct;18(4):4278-4287. doi: 10.3892/ol.2019.10780. Epub 2019 Aug 23.
Diosgenin is an important basic raw material for the production of steroid hormone drugs. It can be isolated and purified from a variety of traditional Chinese medicines or plants. Modern molecular biological studies have shown that diosgenin inhibits various tumor cells migration and invasion ability to varying degrees and . The aim of the present study was to observe the inhibitory effects of diosgenin on the invasive and metastatic capabilities of osteosarcoma cells and to determine the association between the effects of diosgenin on the epithelial-mesenchymal transition (EMT). Wound healing and Transwell assays were used to observe the inhibitory effects of diosgenin on the invasion and migration of two osteosarcoma cell lines. Immunofluorescence was used to observe changes in transforming growth factor β1 (TGF-β1) protein expression levels in the osteosarcoma cells following drug administration. EMT-associated proteins, including TGFβ1, E-cadherin and vimentin were detected by western blotting, which demonstrated that the drug may inhibit the initiation of EMT in osteosarcoma cells. Western blot analysis of the expression of all the proteins in the mitogen-activated protein kinase (MAPK) pathway demonstrated that the drug inhibited the MAPK signaling pathway. The primary mechanism of action of diosgenin was the inhibition of the phosphorylated p38 (pP38) protein. Through a combination of inhibitors of the p38MAPK signaling pathway and detection of the downstream EMT marker protein E-cadherin by quantitative PCR, pP38 was confirmed to be a target of diosgenin in the inhibition of EMT in the osteosarcoma cells via the MAPK molecular signaling pathway. Diosgenin may exhibit utility as an auxiliary drug for the clinical reduction of metastasis in patients with osteosarcoma.
薯蓣皂苷元是生产甾体激素药物的重要基础原料。它可以从多种中药材或植物中分离纯化得到。现代分子生物学研究表明,薯蓣皂苷元能不同程度地抑制多种肿瘤细胞的迁移和侵袭能力。本研究的目的是观察薯蓣皂苷元对骨肉瘤细胞侵袭和转移能力的抑制作用,并确定其对上皮-间质转化(EMT)影响之间的关联。采用伤口愈合实验和Transwell实验观察薯蓣皂苷元对两种骨肉瘤细胞系侵袭和迁移的抑制作用。采用免疫荧光法观察给药后骨肉瘤细胞中转化生长因子β1(TGF-β1)蛋白表达水平的变化。通过蛋白质印迹法检测EMT相关蛋白,包括TGFβ1、E-钙黏蛋白和波形蛋白,结果表明该药物可能抑制骨肉瘤细胞中EMT的启动。对丝裂原活化蛋白激酶(MAPK)信号通路中所有蛋白的表达进行蛋白质印迹分析表明,该药物抑制了MAPK信号通路。薯蓣皂苷元的主要作用机制是抑制磷酸化的p38(pP38)蛋白。通过联合使用p38MAPK信号通路抑制剂,并通过定量PCR检测下游EMT标志物蛋白E-钙黏蛋白,证实pP38是薯蓣皂苷元通过MAPK分子信号通路抑制骨肉瘤细胞EMT的靶点。薯蓣皂苷元可能作为辅助药物用于临床降低骨肉瘤患者的转移风险。