Department of Orthopedic Surgery, Chung Shan Medical University Hospital, Taichung, Taiwan.
Food Chem Toxicol. 2013 Feb;52:193-9. doi: 10.1016/j.fct.2012.11.019. Epub 2012 Nov 28.
Phyllanthus urinaria is widely used as anti-inflammatory, antiviral, antibacterial, and anti-hepatotoxic medicines in almost every tropical country. However, scientific evidence supporting its use in cancer metastasis is limited, particularly osteosarcoma. We investigated the effect of P. urinaria extract (PUE) on cell viability, invasion, and migration in the human osteosarcoma Saos-2 cell line, and looked at the impact of PUE on several relevant proteases and signaling pathways. This study demonstrates that PUE, at a range of concentrations (from 0 to 100 μg/ml), concentration-dependently inhibited the migration/invasion capacities of Saos-2 without cytotoxic effects. Zymographic and western blot analyses revealed that PUE inhibited the urokinase-type plasminogen activator (u-PA) and matrix metalloproteinase-2 (MMP-2) enzyme activity, as well as protein expression. Western blot analysis also showed that PUE inhibits phosphorylation of ERK1/2 and Akt. Testing of mRNA level, quantitative real-time PCR, and promoter assays evaluated the inhibitory effects of PUE on u-PA expression in Saos-2 cells. The chromatin immunoprecipitation (ChIP) assay was reactive to the transcription protein SP-1, which was inhibited by PUE. In conclusion, PUE suppresses human osteosarcoma Saos-2 cell invasion and migration by transcriptionally inhibiting u-PA via ERK and Akt signaling pathways. Therefore, PUE produces anti-metastatic activity in Saos-2 cells.
水飞蓟素在几乎所有的热带国家都被广泛用作抗炎、抗病毒、抗菌和抗肝毒性药物。然而，支持其用于癌症转移的科学证据有限，特别是骨肉瘤。我们研究了水飞蓟素提取物（PUE）对人骨肉瘤 Saos-2 细胞系细胞活力、侵袭和迁移的影响，并观察了 PUE 对几种相关蛋白酶和信号通路的影响。这项研究表明，PUE 在一定浓度范围内（0-100μg/ml），呈浓度依赖性地抑制了 Saos-2 的迁移/侵袭能力，而没有细胞毒性作用。明胶酶谱和 Western blot 分析显示，PUE 抑制了尿激酶型纤溶酶原激活物（u-PA）和基质金属蛋白酶-2（MMP-2）的酶活性和蛋白表达。Western blot 分析还表明，PUE 抑制 ERK1/2 和 Akt 的磷酸化。通过 Saos-2 细胞中 u-PA 表达的 mRNA 水平、定量实时 PCR 和启动子测定测试了 PUE 的抑制作用。染色质免疫沉淀（ChIP）试验对转录蛋白 SP-1 有反应，PUE 抑制了 SP-1。总之，PUE 通过 ERK 和 Akt 信号通路转录抑制 u-PA 抑制人骨肉瘤 Saos-2 细胞的侵袭和迁移。因此，PUE 在 Saos-2 细胞中产生抗转移活性。
