红景天甙诱导骨肉瘤细胞系凋亡并抑制上皮细胞-间充质转化

Salidroside induces apoptosis and inhibits epithelial-mesenchymal transition of human osteosarcoma cell lines

目的探讨红景天甙(Sal)对骨肉瘤细胞系(HOS、U2OS)增殖、凋亡以及上皮细胞-间充质转化(EMT)的影响。方法体外培养骨肉瘤细胞系HOS以及U2OS细胞,用不同浓度Sal处理细胞24 h后,MTT法和流式细胞计量术分别检测细胞活性以及细胞凋亡水平;Western blot检测细胞凋亡蛋白Bcl-2/Bak以及EMT相关分子E-cadherin、N-cadherin、Slug和Snial的表达水平。随后,通过Transwell小室法以及细胞划痕实验检测Sal对细胞侵袭以及迁移的影响。PI3K/Akt抑制剂处理细胞后,Western blot检测EMT相关分子E-cadherin、N-cadherin、Slug和Snial的变化情况。结果不同浓度Sal均能显著抑制HOS及U2OS细胞的细胞活性,并增加细胞凋亡率,且细胞中Bcl-2表达降低,Bak表达增高。150μmol/L的Sal能够显著降低HOS和U2OS细胞的侵袭率以及迁移水平。Western blot结果显示,150μmol/L Sal处理后,HOS和U2OS细胞中E-cadherin表达上调,N-cadherin、Slug和Snial表达下调。PI3K/Akt抑制剂处理后,E-cadherin表达水平较Sal单独处理的细胞进一步升高,而N-cadherin、Slug和Snial表达则显著下降(P<0.05)。结论Sal能够抑制骨肉瘤细胞系细胞的增殖、侵袭以及迁移,并促进细胞凋亡。同时,还能够通过抑制PI3K/Akt信号通路抑制骨肉瘤细胞系发生EMT。

Objective To investigate the effects of salidroside(Sal)on proliferation,apoptosis and epithelial-mesenchymal transition(EMT)of osteosarcoma cell lines(HOS and U2OS).Methods Osteosarcoma HOS and U2OS cells were incubated with different concentrations of Sal for 24 h.Cell activity and apoptosis were detected by MTT assay and flow-cytometry,respectively.Western blot was used to detect the expression of Bcl-2/Bak and EMT-related molecules E-cadherin,N-cadherin,Slug and Snial.Subsequently,the effects of Sal on cell invasion and migration were detected by Transwell assay and cell scratch assay.Finally,the level of EMT-related molecules E-cadherin,N-cadherin,Slug and Snial was detected by Western blot after treatment with PI3K/Akt inhibitor.Results Different concentrations of Sal significantly inhibited cell activity of HOS and U2OS cells,increased the cellular apoptosis rate and expression of Bak,while reduced the expression of Bcl-2 in cells.The invasion rate and migration of HOS and U2OS cells were significantly reduced after incubation with Sal at 150μmol/L.Theexpression of E-cadherin was up-regulated,while N-cadherin,Slug and Snial expression were down-regulated in HOS and U2OS cells after incubation with Sal at 150μmol/L.After incubation with PI3K/Akt inhibitor,the expression of E-cadherin was further increased as compared with the cells treated with Sal alone,while the expression of N-cadherin,Slug and Snial was significantly decreased(P<0.05).Conclusions Sal may inhibit the proliferation,invasion,migration of osteosarcoma cell lines,promote their apoptosis and decreases the EMT through inhiting PI3K/Akt signaling pathway of osteosarcoma cell lines.