曲古抑霉素A对骨肉瘤细胞增殖和迁移能力的影响及其机制

Effect of trichostatin A on the proliferation and migration of osteosarcoma cell lines and its mechanism

目的探讨曲古抑霉素A(TSA)对骨肉瘤细胞增殖和迁移能力的影响及其可能的作用机制。方法不同浓度的TSA处理骨肉瘤MG-63、U2OS细胞48 h,镜下观察细胞形态的改变,CCK8法检测不同浓度对肿瘤细胞的抑制效果,并计算其半数抑制浓度;TSA处理48 h后,以流式细胞术检测对骨肉瘤细胞周期的影响;Transwell法检测TSA对骨肉瘤细胞迁移能力的影响;Western blot检测TSA对骨肉瘤细胞GSK3β、pGSK3β蛋白的影响。结果 TSA处理后,骨肉瘤细胞细胞质减少,失去细胞连接;不同细胞系对TSA的敏感程度不同,与MG-63细胞相比,U2OS细胞更加敏感。流式细胞技术检测结果显示TSA可以使细胞G2/M期增加,G1期和S期减少,将细胞阻滞于G2/M期(P<0.05)。Transwell结果显示TSA可以明显降低骨肉瘤细胞的迁移能力(P<0.05)。Western blot结果表明,TSA并不改变GSK3β的表达,但可以降低pGSK3β的表达(P<0.05)。结论 TSA可以明显抑制骨肉瘤细胞的增殖以及迁移能力,具有显著的抗肿瘤作用,并且这一作用可能是通过下调pGSK3β蛋白表达发挥作用的。

Objective To investigate the effect of trichostatin A （TSA） on proliferation and migration of osteosarcoma and its possible mechanism. Methods Osteosarcoma MG-63 and U2OS cells were treated in different concentrations of TSA for 48 hours. The morphological changes of osteosarcoma were observed under microscope. Calculate the 50%inhibitory concentration （IC50）. After treatment for 48 hours, observe the cell cycle change by flow cytometer;Transwell assay was used to detect the influence of TSA on migration of osteosarcoma;Western blot was used to observe the change of GSK3β, pGSK3β proteins. Results After TSA treatment, cytoplasm was reduced and osteosarcoma lost cell junctions. Different cell lines had different sensitivities to TSA. Compared with the MG-63 cell line, U2OS cell line was more sensitive. Flow cytometry showed that TSA could make G2/M phase increase and G1 phase and S phase decrease, so cells were arrested at G2/M phase. Transwell assay showed TSA could significantly reduce migration of osteosarcoma in vitro. Western blot showed that TSA did not change GSK3βexpression, but it could reduce pGSK3βexpression. Conclusion TSA could inhibit cell proliferation and migration of osteosarcoma. Therefore TSA has significant anti-tumor effect, and this effect may be through down pGSK3βprotein play a role.