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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(6.25~100μmol/L)的Dgg处理骨肉瘤Saos-2细胞96 h后,采用CCK-8法检测细胞增殖情况。用不同浓度(12.5~50μmol/L)的Dgg作用骨肉瘤Saos-2细胞24 h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-B kinase,p-IKK)、磷酸化核因子κB抑制蛋白α(phospho-inhibitor of nuclear factor kappa-Bα,p-IκBα)、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 observe the effect of diosgeninglucoside (Dgg) on proliferation, apoptosis, migration and invasion of human osteosarcoma Saos-2 cells and to explore its possible mechanism. Methods: Osteosarcoma Saos-2 cells were treated with different concentrations of Dgg (6.25 ~ 100μmol / L) for 96 h. Cell proliferation was detected by CCK-8 assay. The osteosarcoma Saos-2 cells were treated with different concentrations of Dgg (12.5-50 μmol / L) for 24 h. The changes of cell apoptosis, migration and invasion were detected by FCM and Transwell chamber assay respectively. Finally, the effect of Dgg (12.5 and 25 μmol / L) on the expression of nuclear factor-κB (NF-κB) / mitogen-activated protein kinase (MAPK) in Saos-2 cells was detected by Western blot ) Pathway related protein and the expression of matrix metalloproteases (MMPs) downstream of this pathway. Results: Dgg could significantly inhibit the proliferation of osteosarcoma Saos-2 cells, induce apoptosis and inhibit cell migration and invasion (all P <0.05) in a concentration-and time-dependent manner. Dgg down-regulated phosphorylation of p38 in NF-|ÊB signaling pathway (P <0.05), but did not affect extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase , JNK) phosphorylation (P values> 0.05). Dgg inhibited phosphorylation of nuclear factor kappa-B kinase (p-IKK), phospho-inhibitor of nuclear factor kappa-B alpha (MAPK) IκBα and phospho-p65 (all P <0.05), and down-regulated the expression of MMP-1, MMP-2 and MMP-9 All <0.05). Conclusion: Dgg can inhibit osteosarcoma cell proliferation, migration and invasion, and promote apoptosis. This effect may be related to tumor cell NF-κB / MAPK signaling pathway is blocked, and MMPs expression is down-regulated.