延龄草皂苷通过NF-κB/MAPK信号通路调控MMPs表达并抑制骨肉瘤Saos-2细胞的迁移和侵袭  

Diosgeninglucoside inhibits migration and invasion of osteosarcoma Saos-2 cells through regulating MMPs expressions via NF-κB/MAPK signaling pathway

作　　者：李文军[1] 康红兵[1] 杨立[1] 聂跃华[1] 艾小红[1] 梅开[1] 

机构地区：[1]南华大学附属第一医院放疗科,湖南衡阳421000

出　　处：《肿瘤》2016年第9期968-976,共9页Tumor

摘　　要：目的：观察延龄草皂苷（diosgeninglucoside，Dgg）对人骨肉瘤Saos-2细胞增殖、凋亡、迁移和侵袭的影响，并探讨其可能的作用机制。方法：应用不同浓度（6．25～100gmol／L）的Dgg处理骨肉瘤Saos-2细胞96h后，采用CCK-8法检测细胞增殖情况。用不同浓度（12．5～50gmol／L）的Dgg作用骨肉瘤Saos-2细胞24h后，分别采用FCM和Transwell小室法检测细胞凋亡、迁移和侵袭能力的变化。最后，采用蛋白质印迹法检测Dgg（12．5和25μmol/L）作用后，Saos-2细胞中核因子κB（nuclear factor—κB，NF-κB）／丝裂原活化蛋白激酶（mitogen—activated protein kinase，MAPK）通路相关蛋白以及该通路下游效应分子基质金属蛋白酶（matrix metalloproteases，MMPs）的表达水平。结果：Dgg可明显抑制骨肉瘤Saos-2细胞的增殖，诱导细胞凋亡，抑制细胞迁移和侵袭（P值均〈0．05），并呈浓度和时间依赖性。Dgg下调NF—κB信号通路中p38的磷酸化（P〈0．05），但不影响细胞外信号调节激酶（extracellular signal—regulated kinase，ERK）和c-Jun氨基末端激酶（c-Jun N—terminal kinase，JNK）的磷酸化（P值均〉0．05）。Dgg抑制MAPK通路中磷酸化核因子κB激酶抑制蛋白（phospho—inhibitor of nuclear factor kappa—Bldnase，p-IKK）、磷酸化核因子κB抑制蛋白α（phospho—inhibitor of nuclear factor kappa—Bα，p-κBα）、IκBα和磷酸化p65（phospho—p65，p-p65）的表达（P值均〈0．05），并下调下游效应分子MMP-1、MMP-2和MMP-9的表达水平（p值均〈0．05）。结论：Dgg可抑制骨肉瘤细胞的增殖、迁移和侵袭，并促进细胞凋亡。这一作用可能与肿瘤细胞中NF—κB／MAPK信号通路被阻滞，以及MMPs表达被下调有关。Objective： To investigate the effects of diosgeninglucoside （Dgg） on proliferation, apoptosis, migration and invasion of human osteosarcoma Saos-2 cells, and to explore their potential molecular mechanisms. Methods： The proliferation of osteosarcoma Saos-2 cells after treatment with different concentrations （6.25-100 μmol/L） of Dgg for 96 h was detected by CCK-8 method. The apoptosis, migration and invasion of Saos-2 cells after treatments with different concentrations （12.5-50 μmol/L） of Dgg for 24 h were detected by FCM and Transwell chamber assay, respectively. Finally, the expressions of nuclear factor-KB/mitogen-activated protein kinase （NF-KB/MAPK） pathway-associated proteins and the downstream effector molecules matrix metalloproteases （MMPs） in Saos-2 cells after Dgg treatment （12.5 and 25μmol/L） were detected by Western blotting. Results： Dgg significantly inhibited the proliferation of Saos-2 cells, induced apoptosis and attenuated cell migration and invasion in a dose- and time-dependent manner （all P 〈 0.05）. Dgg down-regulated the phosphorylation of p38 （P 〈 0.05） in NF-κB signaling pathway, but did not affect the phosphorylated expressions of extracellular signal-regulated kinase （ERK） and c-Jun N-terminal kinase （iNK） （both P 〉 0.05）. Dgg inhibited the expressions of phospho-inhibitor of nuclear factor kappa-B kinase （p-IKK）, phospho-inhibitor of nuclear factor kappa-Bα （p-IκBα）, IκBα and phospho-p65 （p-p65） in MAPK signaling pathway （all P 〈 0.05）. Furthermore, Dgg down-regulated the expressions of MMP-1, MMP-2 and MMP-9 （all P 〈 0.05）. Conclusion： Dgg can inhibit proliferation, migration and invasion of human osteosarcoma Saos-2 cells, and promote apoptosis. These effects may be correlated with down-regulating the expressions of MMPs and inhibiting the activation of NF-~B/MAPK signaling pathway via Dgg treatment.

关 键 词：骨肉瘤 抗肿瘤药(中药) MAP激酶信号系统 基质金属蛋白酶类 细胞迁移分析 肿瘤浸润 延龄草皂苷 

分 类 号：R738.1[医药卫生—肿瘤]