背景:转移导致骨肉瘤5年生存率下降。因此,开发新的骨肉瘤靶向治疗方法势在必行.二氢杨梅素(DHM)具有多种生理功能:它可以抵抗炎症,氧化,和抗肿瘤特性。然而,DHM对骨肉瘤的影响及其潜在机制尚不清楚。
目的:在本研究中,我们研究了DHM在人骨肉瘤U-2OS和HOS细胞中的抗转移特性。
方法:DHM(0、25、50、75和100μM)对细胞活力的影响,迁移,并对入侵进行了检查。西方印迹,RT-PCR,实时定量PCR(QPCR)检测尿激酶型纤溶酶原激活物(uPA)的表达。免疫荧光法检测转录因子SP-1和NF-κB的表达,染色质免疫沉淀测定,和定点诱变荧光素酶报告基因。
结果:我们观察到DHM抑制骨肉瘤细胞系中的细胞迁移和侵袭。此外,DHM通过下调尿激酶纤溶酶原激活物(uPA)表达抑制转移。此外,实时聚合酶链反应和启动子活性测定显示DHM在转录水平降低uPA表达。此外,uPA表达的抑制与SP-1和NF-κB的抑制有关,与uPA启动子结合。无论阻断或诱导细胞外信号调节激酶(ERK)途径,我们证实DHM相关的uPA抑制和细胞转移是通过p-ERK途径发生的.
结论:我们首次提出DHM通过ERK途径抑制骨肉瘤转移,并通过抑制SP-1和NF-κB来抑制下游uPA表达。DHM是针对骨肉瘤的抗转移治疗的潜在治疗剂。
BACKGROUND: Metastasis caused a decline in the 5-years survival rate of osteosarcoma. Therefore, developing new targeted therapeutics for osteosarcoma treatment is imperative. Dihydromyricetin (DHM) has several physiological functions: it counteracts inflammation, oxidation, and antitumor properties. However, the effects of DHM on osteosarcoma and its underlying mechanisms are still not well understood.
OBJECTIVE: In this study, we investigated the antimetastatic properties of DHM in human osteosarcoma U-2 OS and HOS cells.
METHODS: The effects of DHM (0, 25, 50, 75, and 100 μM) on cell viability, migration, and invasion were examined. Western blotting, RT-PCR, and quantitative real-time PCR (QPCR) were determined urokinase plasminogen activator (uPA) expression. The expression of transcriptional factor SP-1 and NF-κB was determined by using immunofluorescence assay, chromatin immunoprecipitation assay, and site-directed mutagenesis luciferase reporter.
RESULTS: We observed that DHM suppresses cell migration and invasion in osteosarcoma cell lines. In addition, DHM inhibits metastasis by downregulating urokinase plasminogen activator (uPA) expression. Moreover, real-time polymerase chain reaction and promoter activity assays revealed that DHM decreased uPA expression at transcription levels. Furthermore, the inhibition of uPA expression was associated with the suppression of SP-1 and NF-κB, which bind to the uPA promoter. Regardless of blocking or inducing the extracellular signal-regulated kinase (ERK) pathway, we verified that the DHM-related suppression of uPA and cell metastasis occurred through the p-ERK pathway.
CONCLUSIONS: We are the first study to propose that DHM suppresses osteosarcoma metastasis through the ERK pathway and through the suppression of SP-1 and NF-κB to inhibit downstream uPA expression. DHM is a potential therapeutic agent for antimetastatic therapy against osteosarcoma.