RhoGDI1通过抑制骨肉瘤细胞凋亡促进其顺铂耐药性的研究

RhoGDI1 promotes cisplatin resistance of osteosarcoma cells by inhibiting apoptosis

目的探讨抑制Rho蛋白鸟苷酸解离抑制因子-1(RhoGDI1)的表达对骨肉瘤顺铂耐药性的影响。方法在骨肉瘤细胞中通过siRNA抑制RhoGDI1的表达(si-RhoGDI1-1、si-RhoGDI1-2组),CCK-8实验检测其对骨肉瘤细胞增殖和顺铂耐药性的影响,划痕实验和体外Transwell检测其对骨肉瘤细胞迁移及侵袭能力的影响,流式细胞术(FACS)和实时荧光定量PCR(RT-qPCR)检测RhoGDI1对骨肉瘤细胞凋亡的影响。结果与si-Control组(转染无意序列)比较,si-RhoGDI1-1、si-RhoGDI1-2组骨肉瘤细胞的增殖速度明细降低(P<0.01),对顺铂的耐受性明显降低(P<0.01),迁移到下室的细胞数显著减少;划痕实验和FACS结果显示,与si-Control组比较,si-RhoGDI1-1、si-RhoGDI1-2组细胞的修复能力明显减弱(P<0.05、P<0.01),细胞凋亡率(P<0.01)和凋亡诱导因子Bax的表达水平明显升高(P<0.01)。结论RhoGDI1通过抑制骨肉瘤细胞凋亡促进骨肉瘤细胞顺铂耐受性。

Objective To investigate the effect of inhibiting the expression of RhoGDI1 on cisplatin resistance of osteosarcoma.Methods The expression of the RhoGDI1 was inhibited by siRNA in the osteosarcoma cells(si-RhoGDI1-1 group,si-RhoGDI1-2 group).The CCK-8 assay was used to determine the effect of RhoGDI1 on the osteosarcoma cell proliferation and the cisplatin resistance of the osteosarcoma cells.The scratch test and the Transwell in vitro were used to detect the effects on the migration and invasion of osteosarcoma cells.The flow cytometry(FACS)and RT-qPCR were used to detect the effect of RhoGDI1 on apoptosis of osteosarcoma cells.Results Compared with the si-Control group(transfected with unintentional sequence),the proliferation rate and cisplatin tolerance of osteosarcoma cells in the si-RhoGDI1-1 and si-RhoGDI1-2 groups decreased significantly(P<0.01),the number of cells migrated to the inferior chamber decreased significantly(P<0.01).The scratch test and FACS results at 12 h and 24 h showed that the cell repair ability of the si-RhoGDI1-1 and si-RhoGDI1-2 groups was significantly weaker than that of the si-Control group(P<0.05,P<0.01).The apoptosis rate and the expression of Bax of the si-RhoGDI1-1 and si-RhoGDI1-2 groups were significantly increased than that of the si-Control group(P<0.01).Conclusion The RhoGDI1 promotes cisplatin resistance of osteosarcoma cells by inhibiting apoptosis.