柴胡皂苷-d促进骨肉瘤细胞凋亡的研究

The study of saikosaponin-d promotes apoptosis of osteosarcoma cells

目的探讨柴胡皂苷-d(SS-d)对骨肉瘤Saos-2细胞增殖、凋亡、迁移和侵袭能力的影响,并探讨其作用机制。方法经不同浓度SS-d处理骨肉瘤Saos-2细胞后,采用八肽胆囊收缩素(CCK-8)法检测骨肉瘤细胞增殖情况,采用流式细胞术检测骨肉瘤细胞凋亡情况,采用Transwell小室法检测骨肉瘤细胞迁移和侵袭能力,最后采用蛋白质免疫印迹(Western blot)法探讨SS-d对核因子-κB(NF-κB)/丝裂原活化蛋白激酶(MAPK)信号转导通路的作用及其下游凋亡相关蛋白的影响。结果 SS-d可呈剂量依赖性地明显抑制骨肉瘤Saos-2细胞的增殖、迁移和侵袭能力(P均<0.05),促进骨肉瘤Saos-2细胞凋亡(P<0.05)。SS-d可抑制p-IκBα和p-p65表达,下调p-Jnk和p-p38表达,进而抑制NF-κB/MAPK信号转导通路激活,促进下游凋亡相关蛋白Bax和裂解活化的半胱氨酸天冬氨酸蛋白酶(cleaved caspase)-3表达,减小Bcl-2/Bax比值,促进骨肉瘤Saos-2细胞凋亡。结论 SS-d可明显抑制骨肉瘤Saos-2细胞增殖、迁移和侵袭,促进骨肉瘤Saos-2细胞凋亡。这一现象与SS-d抑制NF-κB/MAPK信号转导通路激活,减小Bcl-2/Bax比值,促进cleaved caspase-3表达有关。

Objective To investigate the effect of saikosaponin-d（SS-d）on the proliferation,apoptosis,migration and invasion of the osteosarcoma Saos-2 cells,and the related mechanism.Methods Osteosarcoma Saos-2 cells were treated with varying concentrations of SS-d.CCK-8 analysis was used to evaluate the proliferation ability of Saos-2 cells;flow cytometry was employed to investigate the apoptosis ability;Transwell Chamber assay was utilized to explore the migration and invasion ability.Next,Western blot was used to assess the effect of SS-d on activation of nuclear factor-κB（NF-κB）/mitogen-activated protein kinase（MAPK）pathways and the downstream apoptosis-related proteins.Results SS-d dose-dependently inhibited osteosarcoma Saos-2 cell proliferation,migration and invasion（P〈0.05）while promoted apoptosis（P〈0.05）.Western blot revealed that SS-d could inhibit the expressions of p-IκBαand p-p65,reduce the production of p-Jnk and p-p38,contributing to the inhibition of activated NF-κB/MAPK signaling,the elevated expressions of apoptosis-related proteins Bax and cleaved caspase-3 with decreased Bcl-2/Bax ratio,and the apoptosis of osteosarcoma Saos-2 cells.Conclusion SS-d was capable of downregulating osteosarcoma Saos-2 cells proliferation,migration and invasion,upregulating osteosarcoma Saos-2 cells apoptosis.These effects might be associated with the inhibition of NF-κB/MAPK signaling,decreased Bcl-2/Bax ratio and increased cleaved caspase-3 by SS-d treatment.