Pan Po-Jung, Tsai Jai-Jen, Liu Yu-Chang
Department of Physical Medicine and Rehabilitation, National Yang-Ming University Hospital, Yilan, Taiwan, R.O.C.
Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan, R.O.C.
Anticancer Res. 2017 Sep;37(9):4911-4918. doi: 10.21873/anticanres.11900.
The study goal was to investigate effect of amentoflavone on nuclear factor-κB (NF-κB)-modulated metastatic mechanism in osteosarcoma U2OS cells. U2OS cells were treated with amentoflavone, NF-κB inhibitor, protein kinase B (PKB or AKT) inhibitor or mitogen-activated protein kinase (MAPK) inhibitor. Change of cell viability, NF-κB activation, expression of metastasis-associated proteins, signal transduction, and cell migration and invasion were evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, NF-κB reporter gene assay, western blotting, and cell migration and invasion assays. The results demonstrated that inhibition of activation of extracellular signal-regulated kinases (ERK) was a key point for suppression of NF-κB-modulated metastatic mechanism. Amentoflavone significantly inhibited NF-κB activation, ERK phosphorylation, expression of metastasis-associated proteins, and cell migration and invasion. Our findings indicate that amentoflavone reduces metastatic potential through suppression of ERK and NF-κB activation in osteosarcoma U2OS cells.
本研究的目的是探讨穗花杉双黄酮对骨肉瘤U2OS细胞中核因子-κB（NF-κB）调控的转移机制的影响。用穗花杉双黄酮、NF-κB抑制剂、蛋白激酶B（PKB或AKT）抑制剂或丝裂原活化蛋白激酶（MAPK）抑制剂处理U2OS细胞。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法、NF-κB报告基因检测法、蛋白质印迹法以及细胞迁移和侵袭实验，评估细胞活力的变化、NF-κB的激活、转移相关蛋白的表达、信号转导以及细胞迁移和侵袭情况。结果表明，抑制细胞外信号调节激酶（ERK）的激活是抑制NF-κB调控的转移机制的关键环节。穗花杉双黄酮显著抑制NF-κB的激活、ERK磷酸化、转移相关蛋白的表达以及细胞迁移和侵袭。我们的研究结果表明，穗花杉双黄酮通过抑制骨肉瘤U2OS细胞中ERK和NF-κB的激活来降低转移潜能。
